CN101791051B - Preparation method of compound microbial feed additive - Google Patents
Preparation method of compound microbial feed additive Download PDFInfo
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- CN101791051B CN101791051B CN2010101225729A CN201010122572A CN101791051B CN 101791051 B CN101791051 B CN 101791051B CN 2010101225729 A CN2010101225729 A CN 2010101225729A CN 201010122572 A CN201010122572 A CN 201010122572A CN 101791051 B CN101791051 B CN 101791051B
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Abstract
The invention relates to a feed additive, in particular to a prepration method of a compound microbial feed additive, which is used for solving the problems that a variety of feed additives have respective different shortcomings in the actual applications. The preparation method comprises the following steps: preparing selenium yeast, iron yeast and zinc yeast which are rich in trace elements, taking vinegar residue, wheat bran, corn flour, compound Daqu, ammonium sulfate and other auxiliary materials as culture media, adding aspergillus niger, aspergillus oryzae, bacillus subtilis, lactobacillus and yeast, producing a multi-bacterial compound enzyme by the solid-state method, and mixing the functional yeast, phytase and the multi-bacterial compound enzyme for producing a biological feed micro-multi-enzyme which has a large number of live bacteria in beneficial bacterial groups, a complete enzyme system of digestive enzymes and non-digestive enzymes needed by animals and high enzyme activity, and is rich in organic trace elements, vitamins, amino acids and unknown growth-promoting factors with functional nutrition for the animals and people; furthermore, the feed additive has the advantages of advanced process, abundant nutrition, abundant bacterial groups, full enzyme spectrum, high activity, greenness, no pollution and the like.
Description
Technical field
The present invention relates to a kind of feed addictive, be specially a kind of preparation method of composite microorganism type forage additives.
Background technology
Along with the raising of expanding economy and people's living standard, the security that safety and sanitation, nuisanceless, green animal products will enjoy consumer's favor, animal food to have better quality and Geng Gao also be the general trend of events institute to.The relation of animal feed and livestock products is very close, and the former is directly connected to the quality of latter's quality, and the quality of animal products quality is directly connected to human body health again.Animal feed transfers the animal products of supplying with human consumption to; Promptly formed the such food chain of animal feed-animal products-human food; Therefore the security of animal feed is directly connected to people's health and lives safety, how to improve utilization rate, the protection environment of feed simultaneously and ensures that lasting, healthy, the stable development of farming and animal husbandry have become the major issue that the mankind press for solution.
At present, domestic in the breeding process of livestock and poultry, the antibiotic feed that contains commonly used is fed; Not only caused weakening of letting animals feed resistance against diseases, the decline of meat egg product quality and meat flavour, and antibiotic improper use; Will in livestock and poultry meat, milk, egg, produce residually, also possibly cause human cancer, all adverse consequences such as teratogenesis; Therefore, development green non-pollution feed addictive is the research direction of feed industry, and wherein fodder enzyme preparation can improve the digestibility and the utilization rate of feed; Improve the production performance of fowl poultry and fish, can reduce the nitrogen in the fowl poultry excreta, the excretion of phosphorus again, protection water body and soil are avoided polluting; Thereby fodder enzyme preparation as one type efficient, have no side effect and " green " feed addictive of environment-friendly type, in 21 century very wide application prospect will be arranged.
Commodity fodder enzyme preparation great majority in the market are with the sold-in of complex enzyme formulation, like the multienzyme that overflows, security personnel's life etc.In general, complex enzyme formulation is more effective than single enzyme preparation, but and does not mean that the enzyme class is the more the better in the complex enzyme formulation.Complex enzyme formulation has two kinds; Majority is formed by several kinds of single enzyme hybrid modulation, and also having a kind of is to utilize microbial fermentation to produce the complex enzyme formulation of plurality of enzymes, and the former cost is higher; Thereby limited its scope of application, the latter's key technology has two: the one, how to screen the bacterial classification that obtains the high yield enzyme; Another is how to select suitable condition of culture to make bacterial classification bring into play best production performance; From actual operating position; Contained bacterial classification is more single in such complex enzyme formulation of selling on the current market; Can only satisfy the effect that part is disease-resistant or promotion digests, can not satisfy complicated and diversified micro-ecological environment in the organism, and efficiency of feed utilization be lower.
In addition; In the existing feed addictive; For a long time; Trace elements such as iron, zinc, copper, manganese all make an addition in the mixed feed with the form of sulfate, and the existence of sulfate radical not only is prone to cause burn into to influence vitamins stability to feed-processing equipment, and even more serious is the health that can influence animal intestinal after a large amount of sulfate radicals is absorbed by animal.
Summary of the invention
The present invention provides a kind of preparation method of composite microorganism type forage additives in order to solve there are above-mentioned different shortcomings separately in existing all kinds of feed addictives in practical application problem.
The present invention adopts following technical scheme to realize: a kind of preparation method of composite microorganism type forage additives may further comprise the steps:
(1) be rich in the preparation of organic functions yeast of trace element:
A, selenium yeast preparation: brewer's yeast is gone down to posterity on the YEPD inclined-plane, and bacterial classification after the acquisition activation in the YEPD fluid nutrient medium for preparing, is cultivated 20～25h as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH4.5; Brewer's yeast after the activation is inoculated (implication of this percentage is the percentage by weight that inoculum concentration accounts for the seed liquor gross weight) with 3～6% bacterium weight; After cultivating 18～25h, add the sodium selenite solution of 3～6mg/L, continue to be cultured to 45～55h; The zymotic fluid that obtains is centrifugal, collect thalline and oven dry, promptly obtain selenium yeast;
B, the preparation of iron yeast: brewer's yeast is gone down to posterity on the YEPD inclined-plane, and bacterial classification after the acquisition activation in the YEPD fluid nutrient medium for preparing, is cultivated 20～25h as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH6.0 is inoculated the brewer's yeast after the activation with 8～12% bacterium weight, behind cultivation 18～25h, add the copperas solution of 700～900mg/L, continues to be cultured to 55～65h; The zymotic fluid that obtains is centrifugal, collect thalline and oven dry, promptly obtain the iron yeast;
C, the preparation of zinc yeast: brewer's yeast is gone down to posterity on the YEPD inclined-plane, and bacterial classification after the acquisition activation in the YEPD fluid nutrient medium for preparing, is cultivated 20～25h as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH6.0 is inoculated the brewer's yeast after the activation with 8～12% bacterium weight, cultivate 18～25h, adds the solution of zinc sulfate of 700～900mg/L, continues to be cultured to 55～65h; The zymotic fluid that obtains is centrifugal, collect thalline and oven dry, promptly obtain the zinc yeast;
(2) preparation of many bacterium complex enzyme
A, raw material mixing
, wheat bran poor with pretreated raw material vinegar, corn flour are by weight (1.5～2.5): (0.8～1.5): (0.8～1.5) batching; In mixed material, add 3～6% compound Daqus and 0.5～1.2% ammonium sulfate that accounts for the raw material gross weight respectively, stir and to inoculate;
The preparation method of said compound Daqu is following:
The processing of a Daqu raw material: the Daqu raw material is mixed, screens, pulverizes, add water then, make that the weight ratio of Daqu raw material and water is 1: 2.5～3; Said Daqu raw material can be processed by following raw materials by weight percent at least: be with shell wheat 40-45%, buckwheat 18-22%, mung bean 35-40%, perhaps adopt the starter-making materials of traditional Daqu: barley, pea, wheat;
The preparation that the b kind is bent: with the following bacterial classification in the test tube slant culture medium: aspergillus niger (Aspergillus niger) 41254, aspergillus niger (Aspergillus niger) UV-11, monascus (Monascus sp.), Mucor racemosus (Mucor racemosus), graceful radiation Mucor (Actinomucor elegans), Rhizopus oryzae (Rhizopus oryzae), saccharomyces cerevisiae (Saccharomyces cerevisiae), candida krusei (Candida krusei), Acetobacter (Acetobacter sp.), bacillus subtilis (Bacillus subtilis), Lactobacillus delbrueckii (Lactobacillus delbrueckiisubsp.bulgaricus) 6100 are respectively got a ring; Insert in the triangular flask that the Daqu raw material is housed respectively and cultivate; The control fermentation temperature is 30 ℃～33 ℃; Time is 44～46h; The pH value is 5～7; And then proceed enlarged culture, obtain kind of a song;
The preparation of c compound Daqu: adopt raw material system bent, insert solid state fermentation equipment with the Daqu raw material by 1: 45～55 weight ratio with the kind that obtains is bent, after fully mixing, the control fermentation temperature remains on 30 ℃～33 ℃, and the pH value is 5～7, and the blow rate required is 50～75m
3/ min when fermentation time reaches 30～33h, goes out song, then the Daqu that obtains is put into vacuum drying chamber, carries out the cryogenic vacuum oven dry, promptly obtains compound Daqu;
B, inoculation
Use the saccharomycete of cultivating 40～50h, cultivate aspergillus niger and the aspergillus oryzae of 20～30h, the nutrient solution of cultivating the bacillus of 5～10h and this 5 strain bacterium of lactic acid bacteria of cultivating 8～15h is as seed liquor; By the saccharomycete inoculum concentration is 2～4%, and the aspergillus niger inoculum concentration is 2～4%, and the bacillus inoculum concentration is 3～5%; The inoculum concentration of lactic acid bacteria is 3～5%; Aspergillus oryzae inoculum concentration 3～6% o'clock inoculation, the compound Daqu that adds when adding spice, the water content of adjustment compost is 50～65%;
C, solid state fermentation are cultivated
Inoculation back adopts solid state fermentation to cultivate, and cultivation temperature is 30～35 ℃, be cultured to 80～95h after, discharging promptly obtains many bacterium complex enzyme;
(3) with the organic functions yeast that is rich in trace element that obtains in the step (1); The many bacterium complex enzyme and the phytase that obtain in selenium yeast, iron yeast, zinc yeast and the step (2) mix by following weight ratio: selenium yeast 0.5～1.5%; Iron yeast 1～3%; Zinc yeast 1～3%, many bacterium complex enzyme 80.5～89.5%, phytase 8～12%; Low-temperature vacuum drying, pulverizing then promptly obtains composite microorganism type forage additives.This feed addictive can add in the animal daily ration according to 1 ‰ weight ratio in use.
The present invention is rich in organic trace element yeast and phytase with what liquid state fermentation was produced; Many bacterium complex enzyme of producing with solid state fermentation mixes; Many to produce the profitable strain viable bacteria; The digestive ferment that animal needs, non-digestive ferment enzyme are complete, and various enzyme activity is high, and are rich in the little multienzyme of biological feedstuff that animal and human's class is had organic trace element, vitamin, amino acid and the unknown somatomedin of function nutrition.This additive is pale brown toner shape, has fermented flavour, does not have corruption, is as good as stink.Crude protein content>=20%, total viable count>=1 * 10
8, selenium yeast>=0.5mg/kg, zinc yeast>=150mg/kg, iron yeast>=150mg/kg; Phytase>=500u/g, cellulase>=1000u/g, zytase>=1200u/g, pectase>=600u/g; Amylase>=1200u/g, acid protease>=1000u/g, 1,4 beta-glucanase>=800u/g.
In the above-mentioned steps (2), more even for ammonium sulfate is mixed with raw material, can be sprayed directly in the raw material earlier with after the low amounts of water dissolving; Described compound Daqu is in yeast making process; All energy metabolism produces the different enzyme that helps the Daqu fermenting raw materials, and performance micropopulation synergy improves the biochemical property that becomes bent; Produce more metabolite, wherein aspergillus niger 41254 metabolism produce cellulase, pectase, tannase; Aspergillus niger UV-11 metabolism produces carbohydrase; The monascus metabolism produces carbohydrase, look bent; The Mucor racemosus metabolism produces protease; The metabolism of graceful radiation Mucor produces carbohydrase; The Rhizopus oryzae metabolism produces saccharogenic amylase; Saccharomyces cerevisiae 1001 metabolism produce ethanol; The candida krusei metabolism produces aroma-producing yeasts; The Acetobacter metabolism produces acetic acid; The bacillus subtilis metabolism produces diastase, protease; Lactobacillus delbrueckii 6100 metabolism produce lactic acid.Above-mentioned bacterial classification all can buy from market.Through enzyme activity determination, the compound Daqu saccharifying power has had significant raising, can fully decompose carbohydrate in the starter-making materials etc., for the growth of microorganism breeding provides energy, can also effectively improve the utilization rate of grain, reduces the production cost of enterprise; By the access of compound strain, constitute with fungus strain, the enzyme system that enriches Daqu, fungus strain enzyme system is abundant in the song, can obtain a large amount of corresponding purpose metabolites; According to being the research that dynamic change is carried out to the fungus strain enzyme that exists in traditional Daqu; And obtain our the suitableeest required purpose metabolite; Select different strain to insert in the starter-making materials artificially; Do not need in Daqu, to add again other enzyme preparations like this; Also simplified simultaneously production process; Make production technology simple, quick, product quality is safe and reliable.
In this step, utilize that vinegar is poor, wheat bran, corn flour and compound Daqu be primary raw material, add compound Daqu and as the auxiliary materials such as ammonium sulfate of nitrogenous source as culture medium, supply to do microbial reproduction and produce enzyme and use; Microorganism used therefor is basic bacterial strain with compound Daqu; Using is the required microorganism of animal; Aspergillus niger, aspergillus oryzae, bacillus subtilis, Bacillus acidi lactici, the saccharomycete that can produce enzyme again are the compatibility bacterial strain, solid state process produce not only have number of viable greatly but also have high enzyme active add bacterium complex enzyme product.Through the fermentation of micropopulation, it is high to cultivate 20 multiple beneficial bacterium viable counts such as generating saccharomycete, bacillus, Bacillus acidi lactici; Zytase, cellulase, amylase, protease, pectase, dextranase vigor height; Simultaneously wherein compositions such as cellulose, hemicellulose are degraded to the materials such as monose, disaccharide, amino acid, vitamin and unknown somatomedin that the animal advantages of easy digesting absorbs, and are converted into mycoprotein.The interpolation of compound Daqu both provided useful microorganism fungus kind, for the growth of microorganism breeding organic nitrogen source was provided again.
In the above-mentioned steps (3); Said phytase be a kind of can be the enzyme of inositol and phosphoric acid with hydrolysis of phytic acid, can phytic acid and matter hydrochlorate be hydrolyzed into inositol and phosphate after adding phytase in the feed, animal absorbs in confession; Thereby improve the utilization rate and the mineralization of skeleton degree of phosphorus; Can practice thrift phosphor resource, reduce feed cost, it is that disclosed method makes in 200610103778.0 the patent application that this phytase can adopt application number at least.
Composite microorganism type forage additives according to the invention is to utilize vinegar to be pickled with grains or in wine to be matrix; Produce fodder yeast and gathering trace element with adding aspergillus niger, aspergillus oryzae, saccharomyces cerevisiae, bacillus subtilis, the many bacterium of lactic acid bacteria ferment altogether on the basis with the Daqu microorganism, through the green composite microorganism type forage additives of biotechnology production.The feed addictive of profitable strain system, biological multienzyme system, multiple UGF, organic trace element, vitamin, mineral matter, amino acid and carrier composite production livestock and poultry and other animal in the product; Make it to produce collaborative mutually and overall balance effect to animal body; Effect through many bacterium of biology complex enzyme and other material; Fully improve the utilization rate of feed, improve absorption and the conversion thereof of animal, to reach the purpose that promotes that animal increases and practice thrift feed feed.Produce additive prepared using vinegar poor, not only turn waste into wealth, also reduced production cost simultaneously, improved the input-output ratio of feed.The more important thing is that crossing abdomen through animal transforms, can produce the pollution-free foods such as egg, chicken, pork that the mankind had nutritive value.It not only can be prevented and cured diseases, improve food conversion ratio and livestock and poultry production performance; Also can reduce content of cholesterol in the animal products; Reduce content of harmful such as hydrogen sulfide, organophosphor in the ight soil; Obviously reduce animal husbandry and produce pollution on the environment, have remarkable economic efficiency, social benefit and ecological benefits.
In a word, the present invention has the following advantages:
1, technology is advanced: liquid submerged fermentation is cultivated the function yeast, and many bacterium combination and cooperation solid state fermentation is produced many bacterium complex enzyme, and high efficient mixed through low temperature drying, ultramicro grinding, is produced product;
2, nutritious: high-quality Yeast protein, protein can reach more than 20%, and bacterial classification can produce abundant B group vitamin and multiple growth factor, promotes growth of animals or poultry;
3, flora is abundant: the present invention is basic bacterium with the SMEM flora of compound Daqu; And add five kinds of probios such as bacillus, lactic acid bacteria, saccharomycete; It is not to use a kind of microorganism; Neither have only a few kinds of specific microorganisms, but the useful microorganism of numerous species is used as a function colony.Therefore, it is compared with general microbial bacterial agent, not only possesses multiple functional advantage, and in the production of itself, has also shown the high-tech level;
4, zymogram is complete, active high: bacterial classification according to the invention adds various enzyme preparations again simultaneously from enteron aisle, producing a large amount of digestive ferments, has improved the activity of enzyme greatly;
5, primary raw material be that vinegar is poor, wheat bran, corn, the source is abundant, and is cheap, turns waste into wealth;
6, be rich in the function yeast that humans and animals is all had important trophism: selenium yeast, zinc yeast, iron yeast; The function yeast of gathering trace element have other form trace-element complexes incomparable advantage, it will have a wide range of applications in feedstuff industry;
7, use product of the present invention, add supporting scientific culture method, cross abdomen through animal and transform, can produce the mankind are had nutritive value, have the pollution-free foods such as egg, chicken, pork of original local flavor.
The specific embodiment
Embodiment 1:
A kind of preparation method of composite microorganism type forage additives may further comprise the steps:
(1) be rich in the preparation of organic functions yeast of trace element:
A, selenium yeast preparation: brewer's yeast is gone down to posterity twice on the YEPD inclined-plane, bacterial classification after the acquisition activation, in the YEPD fluid nutrient medium for preparing, 28 ℃ of shaken cultivation 20h are as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH4.5 is inoculated the brewer's yeast after the activation with 3% bacterium weight, after 30 ℃ of shaking table reciprocating vibrations are cultivated 18h, add the sodium selenite solution of 3mg/L, continues to be cultured to 45h; The zymotic fluid that obtains at the centrifugal 10min of 4000r/min, is collected thalline and oven dry, promptly obtain selenium yeast;
B, the preparation of iron yeast: brewer's yeast is gone down to posterity on the YEPD inclined-plane, bacterial classification after the acquisition activation, in the YEPD fluid nutrient medium for preparing, 28 ℃ of shaken cultivation 20h are as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH6.0 is inoculated the brewer's yeast after the activation with 8% bacterium weight, after 30 ℃ of shaking table reciprocating vibrations are cultivated 18h, add the copperas solution of people 700mg/L, continues to be cultured to 55h; The zymotic fluid that just obtains is collected thalline and oven dry at the centrifugal 10min of 4000r/min, promptly obtains the iron yeast;
C, the preparation of zinc yeast: brewer's yeast is gone down to posterity on the YEPD inclined-plane, bacterial classification after the acquisition activation, in the YEPD fluid nutrient medium for preparing, 28 ℃ of shaken cultivation 20h are as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH6.0 is inoculated the brewer's yeast after the activation with 8% bacterium weight, 30 ℃ of shaking table reciprocating vibrations are cultivated 18h, add the solution of zinc sulfate of 700mg/L, continue to be cultured to 55h; The zymotic fluid that obtains at the centrifugal 10min of 4000r/min, is collected thalline and oven dry, promptly obtain the zinc yeast;
(2) preparation of many bacterium complex enzyme
A, raw material mixing
, wheat bran poor with pretreated raw material vinegar, corn flour add 3% compound Daqu and 0.5% ammonium sulfate that accounts for the raw material gross weight respectively by weight 1.5: 0.8: 0.8 batchings in mixed material, stir and can inoculate;
The preliminary treatment of raw material comprises: the removal of impurities of raw material, smash, wherein removal step is following: usually with screening and magnetic separation.At first selecting for use receiving sieve to remove has big impurity, silt particle and part dust in the raw material; Then use vibratory sieve, different according to feed particles and impurity size make its automatic classification; When reducing the impurity grain content to greatest extent, remove impurity as much as possible with assurance.Magnetic separation be with the permanent magnetism horseshoe remove that magnetic impurity such as iron in the raw material is cut, iron nail, nut etc.The pulverizing of raw material mainly is crushing maize and breaks up that vinegar is poor, the conglomeration of wheat bran.Make that starch molecule fully contacts with microorganism in the raw material, improve the starch conversion ratio.Use pulverizer, hit down brokenly at the sheet of beating of rotation at a high speed raw material, thin material is by outside the sieve plate hole discharge machine, and the coarse grain continuation is beaten sheet and impacted, up to fragmentation by sieve aperture discharge machine outside, corn flour grinding particle size 1.5～2.0mm.
The preparation method of said compound Daqu is following:
The processing of a Daqu raw material: the Daqu raw material is mixed, screens, pulverizes, add water then, make that the weight ratio of Daqu raw material and water is 1: 2.5;
The preparation that the b kind is bent: with the following bacterial classification in the test tube slant culture medium: aspergillus niger (Aspergillus niger) 41254, aspergillus niger (Aspergillus niger) UV-11, monascus (Monascus sp.), Mucor racemosus (Mucor racemosus), graceful radiation Mucor (Actinomucor elegans), Rhizopus oryzae (Rhizopus oryzae), saccharomyces cerevisiae (Saccharomyces cerevisiae), candida krusei (Candida krusei), Acetobacter (Acetobacter sp.), bacillus subtilis (Bacillus subtilis), Lactobacillus delbrueckii (Lactobacillus delbrueckii subsp.bulgaricus) 6100 are respectively got a ring; Insert in the triangular flask that the Daqu raw material is housed respectively and cultivate; The control fermentation temperature is 30 ℃; Time is 44h; The pH value is 5; And then proceed enlarged culture, obtain kind of a song;
The preparation of c compound Daqu: adopt raw material system bent, with kind song that obtains and the weight ratio access solid state fermentation equipment of Daqu raw material by 1: 45, after fully mixing, the control fermentation temperature remains on 30 ℃, and the pH value is 5, and the blow rate required is 50m
3/ min when fermentation time reaches 30h, goes out song, then the Daqu that obtains is put into vacuum drying chamber, carries out the cryogenic vacuum oven dry, promptly obtains compound Daqu of the present invention; Above-mentioned bacterial classification is existing known products.
B, inoculation
Use the saccharomycete of cultivating 40h, cultivate aspergillus niger and the aspergillus oryzae of 20h, the nutrient solution of cultivating the bacillus of 5h and this 5 strain bacterium of lactic acid bacteria of cultivating 8h is as seed liquor; By the saccharomycete inoculum concentration is 2%, and the aspergillus niger inoculum concentration is 2%, and the bacillus inoculum concentration is 3%; The inoculum concentration of lactic acid bacteria is 3%; Inoculation during aspergillus oryzae inoculum concentration 3%, the compound Daqu that adds when adding spice, the water content of adjustment compost is 50%;
C, solid state fermentation are cultivated
Inoculation back adopts solid state fermentation to cultivate, and cultivation temperature is 30 ℃, be cultured to 80～95h after, discharging promptly obtains many bacterium complex enzyme; Concrete steps are following:
The thickness in raw material dress pond is generally about 30cm, and in order to keep even and good ventilation wind condition, bed of material accumulation should be loosened and be smooth.Fermentation utilizes the ventilation blower air supply and regulates temperature, impels each bacterial classification ramp breeding.Air quantity (m3/h) doubly calculates with the 4-5 that contains total raw material amount (Kg) in the Quchi.And select the model of ventilation blower in view of the above.
Leave standstill and cultivate about 6h, this moment, bent material just began to heat up, and is when temperature rises to 35 left and right sides, just aeration-cooling.After this according to the actual conditions continuous ventilating, material layer temperature is remained on about 30 ℃, the adjusting of temperature can be taked circulating ventilation or control with the mode of ventilation, and the temperature difference of the upper and lower is reduced as far as possible.
In process of production, about 10h, it is very fast to expect that temperature rises after the inoculation, and this moment, song expected that owing to mycelial growth lumps, the resistance of ventilation increases along with the prolongation of mycelial growth time gradually, low and high phenomenon under the appearance of material temperature, and the temperature difference also increases gradually.At this moment, should turn over song with yeast machine immediately, make bent material loose, reduce flowing resistance, keep normal temperature.The interrupted cultivation phenomenons such as contraction fissure occur according to warm rising situation of material and bent material, turns over song again, and the bent material of scarifying is eliminated the crack, in case leak out.Keep about 30 ℃ cultivations of material temperature with continued, be cultured to 80～95h, get final product discharging.Mainly take all factors into consideration that to reach its peak with prolease activity and yeast count be foundation the optimum time of discharging.Selecting 80～95h is the product discharging time.Time is too short, and enzyme activity is not enough, and bacterial classification is fully breeding not; Overlong time, enzyme activity descend on the contrary gradually, and viable count also descends; Influence the Zymolysis Equipment turnover simultaneously, increase the consumption of ventilation blower electric power, increase production cost.
(3) with the organic functions yeast that is rich in trace element that obtains in the step (1); The many bacterium complex enzyme and the phytase that obtain in selenium yeast, iron yeast, zinc yeast and the step (2) mix by following weight ratio: selenium yeast 0.5%, iron yeast 1%, zinc yeast 1%; Many bacterium complex enzyme 89.5%; Phytase 8% fully mixes 10～20min, till mixing in horizontal spiral ribbon mixer; Low-temperature vacuum drying then, dry technological parameter is: 30 ℃ of temperature, when vacuum was 0.06-0.096Mpa, be 3～4h drying time, product water content≤13%; Dried material is pulverized with pulverizer, pulverizes the satisfactory material of fineness of finished product, promptly obtains composite microorganism type forage additives of the present invention.
Embodiment 2:
A kind of preparation method of composite microorganism type forage additives may further comprise the steps:
(1) be rich in the preparation of organic functions yeast of trace element:
A, selenium yeast preparation: brewer's yeast is gone down to posterity twice on the YEPD inclined-plane, bacterial classification after the acquisition activation, in the YEPD fluid nutrient medium for preparing, 28 ℃ of shaken cultivation 25h are as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH4.5 is inoculated the brewer's yeast after the activation with 6% bacterium weight, after 30 ℃ of shaking table reciprocating vibrations are cultivated 25h, add the sodium selenite solution of 6mg/L, continues to be cultured to 55h; The zymotic fluid that obtains at the centrifugal 10min of 4000r/min, is collected thalline and oven dry, promptly obtain selenium yeast;
B, the preparation of iron yeast: brewer's yeast is gone down to posterity on the YEPD inclined-plane, bacterial classification after the acquisition activation, in the YEPD fluid nutrient medium for preparing, 28 ℃ of shaken cultivation 25h are as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH6.0 is inoculated the brewer's yeast after the activation with 12% bacterium weight, after 30 ℃ of shaking table reciprocating vibrations are cultivated 25h, add the copperas solution of 900mg/L, continues to be cultured to 65h; The zymotic fluid that just obtains is collected thalline and oven dry at the centrifugal 10min of 4000r/min, promptly obtains the iron yeast;
C, the preparation of zinc yeast: brewer's yeast is gone down to posterity on the YEPD inclined-plane, bacterial classification after the acquisition activation, in the YEPD fluid nutrient medium for preparing, 28 ℃ of shaken cultivation 25h are as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH6.0 is inoculated the brewer's yeast after the activation with 12% bacterium weight, 30 ℃ of shaking table reciprocating vibrations are cultivated 25h, add the solution of zinc sulfate of 900mg/L, continue to be cultured to 65h; The zymotic fluid that obtains at the centrifugal 10min of 4000r/min, is collected thalline and oven dry, promptly obtain the zinc yeast;
(2) preparation of many bacterium complex enzyme
A, raw material mixing
, wheat bran poor with pretreated raw material vinegar, corn flour add 6% compound Daqu and 1.2% ammonium sulfate that accounts for the raw material gross weight respectively by weight 2.5: 1.5: 1.5 batchings in mixed material, stir and can inoculate;
The preliminary treatment of raw material comprises: the removal of impurities of raw material, smash, wherein removal step is following: usually with screening and magnetic separation.At first selecting for use receiving sieve to remove has big impurity, silt particle and part dust in the raw material; Then use vibratory sieve, different according to feed particles and impurity size make its automatic classification; When reducing the impurity grain content to greatest extent, remove impurity as much as possible with assurance.Magnetic separation be with the permanent magnetism horseshoe remove that magnetic impurity such as iron in the raw material is cut, iron nail, nut etc.The pulverizing of raw material mainly is crushing maize and breaks up that vinegar is poor, the conglomeration of wheat bran.Make that starch molecule fully contacts with microorganism in the raw material, improve the starch conversion ratio.Use pulverizer, hit down brokenly at the sheet of beating of rotation at a high speed raw material, thin material is by outside the sieve plate hole discharge machine, and the coarse grain continuation is beaten sheet and impacted, up to fragmentation by sieve aperture discharge machine outside, corn flour grinding particle size 1.5～2.0mm.
The preparation method of said compound Daqu is following:
The processing of a Daqu raw material: the Daqu raw material is mixed, screens, pulverizes, add water then, make that the weight ratio of Daqu raw material and water is 1: 3;
The preparation that the b kind is bent: with the following bacterial classification in the test tube slant culture medium: aspergillus niger (Aspergillus niger) 41254, aspergillus niger (Aspergillus niger) UV-11, monascus (Monascus sp.), Mucor racemosus (Mucor racemosus), graceful radiation Mucor (Actinomucor elegans), Rhizopus oryzae (Rhizopus oryzae), saccharomyces cerevisiae (Saccharomyces cerevisiae), candida krusei (Candida kmsei), Acetobacter (Acetobacter sp.), bacillus subtilis (Bacillus subtilis), Lactobacillus delbrueckii (Lactobacillus delbrueckii subsp.bulgaricus) 6100 are respectively got a ring; Insert in the triangular flask that the Daqu raw material is housed respectively and cultivate; The control fermentation temperature is 33 ℃; Time is 46h; The pH value is 7; And then proceed enlarged culture, obtain kind of a song;
The preparation of c compound Daqu: adopt raw material system bent, with kind song that obtains and the weight ratio access solid state fermentation equipment of Daqu raw material by 1: 55, after fully mixing, the control fermentation temperature remains on 33 ℃, and the pH value is 7, and the blow rate required is 75m
3/ min when fermentation time reaches 33h, goes out song, then the Daqu that obtains is put into vacuum drying chamber, carries out the cryogenic vacuum oven dry, promptly obtains compound Daqu of the present invention; Above-mentioned bacterial classification is existing known products.
B, inoculation
Use the saccharomycete of cultivating 40～50h, cultivate aspergillus niger and the aspergillus oryzae of 30h, the nutrient solution of cultivating the bacillus of 10h and this 5 strain bacterium of lactic acid bacteria of cultivating 15h is as seed liquor; By the saccharomycete inoculum concentration is 4%, and the aspergillus niger inoculum concentration is 4%, and the bacillus inoculum concentration is 5%; The inoculum concentration of lactic acid bacteria is 5%; Inoculation during aspergillus oryzae inoculum concentration 6%, the compound Daqu that adds when adding spice, the water content of adjustment compost is 65%;
C, solid state fermentation are cultivated
Inoculation back adopts solid state fermentation to cultivate, and cultivation temperature is 35 ℃, be cultured to 80～95h after, discharging promptly obtains many bacterium complex enzyme; Concrete steps are following:
The thickness in raw material dress pond is generally about 30cm, and in order to keep even and good ventilation wind condition, bed of material accumulation should be loosened and be smooth.Fermentation utilizes the ventilation blower air supply and regulates temperature, impels each bacterial classification ramp breeding.Air quantity (m
3/ h) doubly calculate with the 4-5 that contains total raw material amount (Kg) in the Quchi.And select the model of ventilation blower in view of the above.
Leave standstill and cultivate about 6h, this moment, bent material just began to heat up, and is when temperature rises to 35 left and right sides, just aeration-cooling.After this according to the actual conditions continuous ventilating, material layer temperature is remained on about 30 ℃, the adjusting of temperature can be taked circulating ventilation or control with the mode of ventilation, and the temperature difference of the upper and lower is reduced as far as possible.
In process of production, about 10h, it is very fast to expect that temperature rises after the inoculation, and this moment, song expected that owing to mycelial growth lumps, the resistance of ventilation increases along with the prolongation of mycelial growth time gradually, low and high phenomenon under the appearance of material temperature, and the temperature difference also increases gradually.At this moment, should turn over song with yeast machine immediately, make bent material loose, reduce flowing resistance, keep normal temperature.The interrupted cultivation phenomenons such as contraction fissure occur according to warm rising situation of material and bent material, turns over song again, and the bent material of scarifying is eliminated the crack, in case leak out.Keep about 30 ℃ cultivations of material temperature with continued, be cultured to 80～95h, get final product discharging.Mainly take all factors into consideration that to reach its peak with prolease activity and yeast count be foundation the optimum time of discharging.Selecting 80～95h is the product discharging time.Time is too short, and enzyme activity is not enough, and bacterial classification is fully breeding not; Overlong time, enzyme activity descend on the contrary gradually, and viable count also descends; Influence the Zymolysis Equipment turnover simultaneously, increase the consumption of ventilation blower electric power, increase production cost.
(3) with the organic functions yeast that is rich in trace element that obtains in the step (1); The many bacterium complex enzyme and the phytase that obtain in selenium yeast, iron yeast, zinc yeast and the step (2) mix by following weight ratio: selenium yeast 1.5%, iron yeast 3%, zinc yeast 3%; Many bacterium complex enzyme 80.5%; Phytase 12% fully mixes 10～20min, till mixing in horizontal spiral ribbon mixer; Low-temperature vacuum drying then, dry technological parameter is: 30 ℃ of temperature, when vacuum was 0.06-0.096Mpa, be 3～4h drying time, product water content≤13%; Dried material is pulverized with pulverizer, pulverizes the satisfactory material of fineness of finished product, promptly obtains composite microorganism type forage additives of the present invention.
Embodiment 3:
A kind of preparation method of composite microorganism type forage additives may further comprise the steps:
(1) be rich in the preparation of organic functions yeast of trace element:
A, selenium yeast preparation: brewer's yeast is gone down to posterity twice on the YEPD inclined-plane, bacterial classification after the acquisition activation, in the YEPD fluid nutrient medium for preparing, 28 ℃ of shaken cultivation 23h are as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH4.5 is inoculated the brewer's yeast after the activation with 5% bacterium weight, after 30 ℃ of shaking table reciprocating vibrations are cultivated 20h, add the sodium selenite solution of 5mg/L, continues to be cultured to 48h; The zymotic fluid that obtains at the centrifugal 10min of 4000r/min, is collected thalline and oven dry, promptly obtain selenium yeast;
B, the preparation of iron yeast: brewer's yeast is gone down to posterity on the YEPD inclined-plane, bacterial classification after the acquisition activation, in the YEPD fluid nutrient medium for preparing, 28 ℃ of shaken cultivation 22h are as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH6.0 is inoculated the brewer's yeast after the activation with 10% bacterium weight, after 30 ℃ of shaking table reciprocating vibrations are cultivated 20h, add the copperas solution of 800mg/L, continues to be cultured to 60h; The zymotic fluid that just obtains is collected thalline and oven dry at the centrifugal 10min of 4000r/min, promptly obtains the iron yeast;
C, the preparation of zinc yeast: brewer's yeast is gone down to posterity on the YEPD inclined-plane, bacterial classification after the acquisition activation, in the YEPD fluid nutrient medium for preparing, 28 ℃ of shaken cultivation 20h are as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH6.0 is inoculated the brewer's yeast after the activation with 10% bacterium weight, 30 ℃ of shaking table reciprocating vibrations are cultivated 20h, add the solution of zinc sulfate of 800mg/L, continue to be cultured to 60h; The zymotic fluid that obtains at the centrifugal 10min of 4000r/min, is collected thalline and oven dry, promptly obtain the zinc yeast;
(2) preparation of many bacterium complex enzyme
A, raw material mixing
, wheat bran poor with pretreated raw material vinegar, corn flour add 5% compound Daqu and 1% ammonium sulfate that accounts for the raw material gross weight respectively by weight 2: 1: 1 batchings in mixed material, stir and can inoculate;
The preliminary treatment of raw material comprises: the removal of impurities of raw material, smash, wherein removal step is following: usually with screening and magnetic separation.At first selecting for use receiving sieve to remove has big impurity, silt particle and part dust in the raw material; Then use vibratory sieve, different according to feed particles and impurity size make its automatic classification; When reducing the impurity grain content to greatest extent, remove impurity as much as possible with assurance.Magnetic separation be with the permanent magnetism horseshoe remove that magnetic impurity such as iron in the raw material is cut, iron nail, nut etc.The pulverizing of raw material mainly is crushing maize and breaks up that vinegar is poor, the conglomeration of wheat bran.Make that starch molecule fully contacts with microorganism in the raw material, improve the starch conversion ratio.Use pulverizer, hit down brokenly at the sheet of beating of rotation at a high speed raw material, thin material is by outside the sieve plate hole discharge machine, and the coarse grain continuation is beaten sheet and impacted, up to fragmentation by sieve aperture discharge machine outside, corn flour grinding particle size 1.5～2.0mm.
The preparation method of said compound Daqu is following:
The processing of a Daqu raw material: the Daqu raw material is mixed, screens, pulverizes, add water then, make that the weight ratio of Daqu raw material and water is 1: 2.8;
The preparation that the b kind is bent: with the following bacterial classification in the test tube slant culture medium: aspergillus niger (Aspergillus niger) 41254, aspergillus niger (Aspergillus niger) UV-11, monascus (Monascus sp.), Mucor racemosus (Mucor racemosus), graceful radiation Mucor (Actinomucor elegans), Rhizopus oryzae (Rhizopus oryzae), saccharomyces cerevisiae (Saccharomyces cerevisiae), candida krusei (Candida krusei), Acetobacter (Acetobacter sp.), bacillus subtilis (Bacillus subtilis), Lactobacillus delbrueckii (Lactobacillus delbrueckii subsp.bulgaricus) 6100 are respectively got a ring; Insert in the triangular flask that the Daqu raw material is housed respectively and cultivate; The control fermentation temperature is 32 ℃; Time is 45h; The pH value is 6; And then proceed enlarged culture, obtain kind of a song;
The preparation of c compound Daqu: adopt raw material system bent, with kind song that obtains and the weight ratio access solid state fermentation equipment of Daqu raw material by 1: 50, after fully mixing, the control fermentation temperature remains on 31 ℃, and the pH value is 6, and the blow rate required is 60m
3/ min when fermentation time reaches 32h, goes out song, then the Daqu that obtains is put into vacuum drying chamber, carries out the cryogenic vacuum oven dry, promptly obtains compound Daqu of the present invention; Above-mentioned bacterial classification is existing known products.
B, inoculation
Use the saccharomycete of cultivating 45h, cultivate aspergillus niger and the aspergillus oryzae of 24h, the nutrient solution of cultivating the bacillus of 8h and this 5 strain bacterium of lactic acid bacteria of cultivating 10h is as seed liquor; By the saccharomycete inoculum concentration is 3%, and the aspergillus niger inoculum concentration is 3%, and the bacillus inoculum concentration is 4%; The inoculum concentration of lactic acid bacteria is 4%; Inoculation during aspergillus oryzae inoculum concentration 4%, the compound Daqu that adds when adding spice, the water content of adjustment compost is 60%;
C, solid state fermentation are cultivated
Inoculation back adopts solid state fermentation to cultivate, and cultivation temperature is 30～35 ℃, be cultured to 80～95h after, discharging promptly obtains many bacterium complex enzyme; Concrete steps are following:
The thickness in raw material dress pond is generally about 30cm, and in order to keep even and good ventilation wind condition, bed of material accumulation should be loosened and be smooth.Fermentation utilizes the ventilation blower air supply and regulates temperature, impels each bacterial classification ramp breeding.Air quantity (m3/h) doubly calculates with the 4-5 that contains total raw material amount (Kg) in the Quchi.And select the model of ventilation blower in view of the above.
Leave standstill and cultivate about 6h, this moment, bent material just began to heat up, and is when temperature rises to 35 left and right sides, just aeration-cooling.After this according to the actual conditions continuous ventilating, material layer temperature is remained on about 30 ℃, the adjusting of temperature can be taked circulating ventilation or control with the mode of ventilation, and the temperature difference of the upper and lower is reduced as far as possible.
In process of production, about 10h, it is very fast to expect that temperature rises after the inoculation, and this moment, song expected that owing to mycelial growth lumps, the resistance of ventilation increases along with the prolongation of mycelial growth time gradually, low and high phenomenon under the appearance of material temperature, and the temperature difference also increases gradually.At this moment, should turn over song with yeast machine immediately, make bent material loose, reduce flowing resistance, keep normal temperature.The interrupted cultivation phenomenons such as contraction fissure occur according to warm rising situation of material and bent material, turns over song again, and the bent material of scarifying is eliminated the crack, in case leak out.Keep about 30 ℃ cultivations of material temperature with continued, be cultured to 80～95h, get final product discharging.Mainly take all factors into consideration that to reach its peak with prolease activity and yeast count be foundation the optimum time of discharging.Selecting 80～95h is the product discharging time.Time is too short, and enzyme activity is not enough, and bacterial classification is fully breeding not; Overlong time, enzyme activity descend on the contrary gradually, and viable count also descends; Influence the Zymolysis Equipment turnover simultaneously, increase the consumption of ventilation blower electric power, increase production cost.
(3) with the organic functions yeast that is rich in trace element that obtains in the step (1); The many bacterium complex enzyme and the phytase that obtain in selenium yeast, iron yeast, zinc yeast and the step (2) mix by following weight ratio: selenium yeast 1%, iron yeast 2%, zinc yeast 2%; Many bacterium complex enzyme 85%; Phytase 10% fully mixes 10～20min, till mixing in horizontal spiral ribbon mixer; Low-temperature vacuum drying then, dry technological parameter is: 30 ℃ of temperature, when vacuum was 0.06-0.096Mpa, be 3～4h drying time, product water content≤13%; Dried material is pulverized with pulverizer, pulverizes the satisfactory material of fineness of finished product, promptly obtains composite microorganism type forage additives of the present invention.
Claims (1)
1. the preparation method of a composite microorganism type forage additives is characterized in that may further comprise the steps:
(1) is rich in the preparation of organic functions yeast of trace element
A, selenium yeast preparation: brewer's yeast is gone down to posterity on the YEPD inclined-plane, and bacterial classification after the acquisition activation in the YEPD fluid nutrient medium for preparing, is cultivated 20～25h as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH4.5 is inoculated the brewer's yeast after the activation with 3～6% bacterium weight, behind cultivation 18～25h, add the sodium selenite solution of 3～6mg/L, continues to be cultured to 45～55h; The zymotic fluid that obtains is centrifugal, collect thalline and oven dry, promptly obtain selenium yeast;
B, the preparation of iron yeast: brewer's yeast is gone down to posterity on the YEPD inclined-plane, and bacterial classification after the acquisition activation in the YEPD fluid nutrient medium for preparing, is cultivated 20～25h as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH6.0 is inoculated the brewer's yeast after the activation with 8～12% bacterium weight, behind cultivation 18～25h, add the copperas solution of 700～900mg/L, continues to be cultured to 55～65h; The zymotic fluid that obtains is centrifugal, collect thalline and oven dry, promptly obtain the iron yeast;
C, the preparation of zinc yeast: brewer's yeast is gone down to posterity on the YEPD inclined-plane, and bacterial classification after the acquisition activation in the YEPD fluid nutrient medium for preparing, is cultivated 20～25h as seed liquor with bacterial classification inoculation after the activation; The YEPD fluid nutrient medium of preparation pH6.0 is inoculated the brewer's yeast after the activation with 8～12% bacterium weight, cultivate 18～25h, adds the solution of zinc sulfate of 700～900mg/L, continues to be cultured to 55～65h; The zymotic fluid that obtains is centrifugal, collect thalline and oven dry, promptly obtain the zinc yeast;
(2) preparation of many bacterium complex enzyme
A, raw material mixing
, wheat bran poor with pretreated raw material vinegar, corn flour are by weight (1.5～2.5): (0.8～1.5): (0.8～1.5) batching; In mixed material, add 3～6% compound Daqus and 0.5～1.2% ammonium sulfate that accounts for the raw material gross weight respectively, stir and to inoculate;
The preparation method of said compound Daqu is following:
The processing of a Daqu raw material: the Daqu raw material is mixed, screens, pulverizes, add water then, make that the weight ratio of Daqu raw material and water is 1: 2.5～3;
The preparation that the b kind is bent: with the following bacterial classification in the test tube slant culture medium: aspergillus niger (Aspergillus niger) 41254, aspergillus niger (Aspergillus niger) UV-11, monascus (Monascus sp.), Mucor racemosus (Mucor racemosus), graceful radiation Mucor (Actinomucor elegans), Rhizopus oryzae (Rhizopus oryzae), saccharomyces cerevisiae (Saccharomyces cerevisiae), candida krusei (Candida krusei), Acetobacter (Acetobacter sp.), bacillus subtilis (Bacillus subtilis), Lactobacillus delbrueckii (Lactobacillus delbrueckiisubsp.bulgaricus) 6100 are respectively got a ring; Insert in the triangular flask that the Daqu raw material is housed respectively and cultivate; The control fermentation temperature is 30 ℃～33 ℃; Time is 44～46h; The pH value is 5～7; And then proceed enlarged culture, obtain kind of a song;
The preparation of c compound Daqu: adopt raw material system bent, insert solid state fermentation equipment with the Daqu raw material by 1: 45～55 weight ratio with the kind that obtains is bent, after fully mixing, the control fermentation temperature remains on 30 ℃～33 ℃, and the pH value is 5～7, and the blow rate required is 50～75m
3/ min when fermentation time reaches 30～33h, goes out song, then the Daqu that obtains is put into vacuum drying chamber, carries out the cryogenic vacuum oven dry, promptly obtains compound Daqu;
B, inoculation
Use the saccharomycete of cultivating 40～50h, cultivate aspergillus niger and the aspergillus oryzae of 20～30h, the nutrient solution of cultivating the bacillus of 5～10h and this 5 strain bacterium of lactic acid bacteria of cultivating 8～15h is as seed liquor; By the saccharomycete inoculum concentration is 2～4%, and the aspergillus niger inoculum concentration is 2～4%, and the bacillus inoculum concentration is 3～5%; The inoculum concentration of lactic acid bacteria is 3～5%; Aspergillus oryzae inoculum concentration 3～6% o'clock inoculation, the compound Daqu that adds when adding spice, the water content of adjustment compost is 50～65%;
C, solid state fermentation are cultivated
Inoculation back adopts solid state fermentation to cultivate, and cultivation temperature is 30～35 ℃, be cultured to 80～95h after, discharging promptly obtains many bacterium complex enzyme;
(3) with the organic functions yeast that is rich in trace element that obtains in the step (1); The many bacterium complex enzyme and the phytase that obtain in selenium yeast, iron yeast, zinc yeast and the step (2) mix by following weight ratio: selenium yeast 0.5～1.5%; Iron yeast 1～3%; Zinc yeast 1～3%, many bacterium complex enzyme 80.5～89.5%, phytase 8～12%; Low-temperature vacuum drying, pulverizing then promptly obtains composite microorganism type forage additives.
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