CN1147581C - Preparation method of complex microecological feed additive richly containing wood oligose - Google Patents
Preparation method of complex microecological feed additive richly containing wood oligoseInfo
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- CN1147581C CN1147581C CNB011274425A CN01127442A CN1147581C CN 1147581 C CN1147581 C CN 1147581C CN B011274425 A CNB011274425 A CN B011274425A CN 01127442 A CN01127442 A CN 01127442A CN 1147581 C CN1147581 C CN 1147581C
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- wood oligose
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- water
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- enzyme liquid
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
Abstract
The present invention relates to a method for preparing composite micro-ecological feed additives richly containing wood oligose, and belongs to the technical field of microbial feed additive application. The present invention adopts aspergillus niger An-76 with high proteinase yield as bacterial strains, takes corn cobs, wheat bran, ammonium sulfate and water as auxiliary materials, and inoculates An76 secondary bacteria. Fermented culture materials are immersed in water. Filtered supernatant fluid is the coarse enzyme liquid of xylanase. The coarse enzyme liquid of xylanase is mixed with corn stalks and/or corn cob raw materials exploded by steam. The weight ratio of the coarse enzyme liquid of xylanase to the corn stalks and/or corn cob raw materials is (10 to 11) to 1. The mixture is filtered after enzymolysis. Supernatant liquid is concentrated in order to obtain the concentrated liquid of wood oligose. The concentrated liquid of wood oligose is adsorbed by endomycopsis fibuligera dekk yeast culture materials, and is dried. The present invention combines enzyme engineering technology with fermentation engineering technology together, and has the dual functions of viable bacterium preparation and double-dismutase factors. The present invention develops and utilizes stalk resources, does not discharge waster water, waster gas and waster slag simultaneously, and avoids serious environment pollution caused by stalk incineration.
Description
(1) technical field
The present invention relates to a kind of preparation method of wood oligose fodder additives, belong to the using microbe field of feed additive technology.
(2) background technology
Keep cultivated animals health, people's common antibiotics kills harmful microorganism, but at this moment, beneficial microorganism also major part is killed.And, in recent years, because antibiotic feed additive exists inevitably some hidden danger, cause animal and human's allergy, teratogenesis, canceration etc. as drug residue, serial problems such as the generation of Resistant strain in addition, destruction animal intestinal microecological balance, therefore many developed countries have forbidden or have limited and use some microbiotic and chemical classes growth stimulant, developing nontoxic, harmless, noresidue, free from environmental pollution, the additive that do not develop immunity to drugs, is to reach the 21st century countries in the world new additive agent of exploitation mutually unexpectedly now.Disease is cured the disease, is the behavior of mechanical medical science, and not sick diseases prevention is biomedical behavior, and anosis health care is the behavior of ecological medical science, and medical science has absorbed ecological viewpoint and just formed ecological medical science.Ecological medical science is the highest developmental stage of modern medicine, and what emphasize is the unification of human body and interior environment, belong to microecology (Microecology) category, and microecological balance (Microeubiosis) is the key problem of microecology.A large amount of microorganism survival and reproduction (about 100,000,000) is arranged in the livestock and poultry alimentary tract, can be divided into profitable strain, pernicious bacteria and marginal facultative flora according to its secretion behavior.Be mutual restriction between the microflora in the digestive tube, when wherein a part of group takes up space advantage in enteron aisle, a part of in addition flora then is in a disadvantageous position, at this moment the population equilibrium of microorganism is just destroyed in the digestive tube, if deflection pernicious bacteria, then can cause the disorder of cultivated animals digestive system function, bring out diarrhea, constipation, hypoevolutism, immunity system disease such as be obstructed.Generally, the livestock and poultry outward appearance be it seems very healthy, and the overwhelming majority is in sub-health state in fact, that is to say, when external conditions changed, the livestock and poultry resistivity reduced, and disease will occur.Therefore,,, utilize normal microflora member or its to promote the appearance of the probiotics that material is made, be historical certainty, and at home and abroad emerge rapidly in recent years in order to adjust microecological balance according to the microecology principle.Probiotics is called Probiotics abroad, and domestic translated name is more, as probiotics, gives birth to rhzomorph, beneficial rhzomorph etc., U.S.'s instrument and drug administration this type of Product Definition be " microbial product of directly feeding " (Direct Fed Microbials, DFM).Its mainly act on be that weightening finish is fattened, the vital role of the trans-utilization rate of disease preventing and treating, raising breeding performonce fo animals and feed.Yeast is used for a quasi-microorganism of " microbial product of directly feeding " exactly.After yeast cell enters digestive tube,, consume the oxygen in the digestive tube, be anaerobic bacteria flora useful in the enteron aisle, lay the foundation as the propagation of bifidus bacillus in digestive tube leading portion propagation.In recent ten years, it is found that genus bifidobacterium plays important effect in cultivated animals digestive tubes such as livestock and poultry, the survival volume of bifidus bacillus is very big to digestion, absorptive function influence, so people have developed multiple bifidobacterium preparations.But because bifidus bacillus is an obligatory anaerobic bacteria, to photo-labile, not acidproof, non-refractory, bring certain difficulty to industrial production, tame out aerotolerant bifidus bacillus although separated in recent years, created good condition for the exploitation active bacteria formulation, but it is through the effect of gastric juice (pH0.9-1.5), intestinal juice (about pH8.3), the viable count that arrives enteron aisle obviously reduces.In addition, bifidus bacillus is very fragile, uses with microbiotic, often is killed, and survival rate is little, thereby DeGrain.Therefore, people are in the research active bacteria formulation, interest has been taken place in inherent microorganism species in the animal body, if can add some artificially in feed can not be utilized by cultivated animals self, can not utilize for the harmful bacterium of major part again, and can only utilize for bifidus bacillus, thereby the material that bifidus bacillus is bred then can get twice the result with half the effort on effect.This class probiotics is called bifidus factor (Bifiaus factor BF).BF is that the promotion bifidus bacillus that a big class formation and character have nothing in common with each other breeds necessary non-toxic substance.Oligosaccharides is a most important class among the BF, as mannooligo saccharide, different wheat tooth oligosaccharides, Nutriflora P, wood oligose, alga oligosaccharide etc.
(3) summary of the invention
The present invention utilizes corn stalk etc. to be rich in the cheap raw material of xylan, the using microbe enzyme engineering technology, the Production by Enzymes wood oligose, adsorb with yeast culture then, make and not only contain two qi factor wood oligoses but also contain the novel probiotics of active bacteria formulation zymic, be applied to livestock and poultry breeding industry, substitute microbiotic, have excellent results.
Content of the present invention comprises that (1) adopts high aspergillus niger (Aspergillus niger) An-76 of proteolytic enzyme productive rate is bacterial strain, be equipped with corn cob, wheat bran, ammonium sulfate and water, inoculation An76 second class inoculum, fermentation, fermenting culture is soaked, and filtering supernatant is the crude enzyme liquid of zytase; (2) the zytase crude enzyme liquid is pressed (10～11) with corn stalk, the core hemicellulose raw material of steam explosion: 1 weight ratio is mixed, and enzymolysis filters, and supernatant concentration obtains the oligosaccharides concentrated solution; (3) oligosaccharides concentrated solution and Endomycopsis Fibnligera yeast culture adsorb at 1: 1, and oven dry is product of the present invention.
The preparation of above-mentioned Endomycopsis Fibnligera yeast culture can be adopted prior art, as disclosed CN1286037A specification sheets page 1 on March 7 calendar year 2001.Can Endomycopsis Fibnligera (Ecomycopsis fibuligea) be bacterial strain also, with 30～40 parts of dregs of beans, 50～60 parts of corns, 20～30 parts in wheat bran, ammonium sulfate be substratum for 1～2 part, raw material mixes through pulverizing, profit water, water content 55～65%, sterilization, connect 5～7 parts of Endomycopsis Fibnligera second class inoculums, shallow-layer fermentation 20～24 hours, 28～30 ℃ of temperature, relative humidity 80～85%, cryodrying below 80 ℃ is pulverized.More than be weight part.
Method of the present invention comprises following concrete steps:
1. the preparation of zytase crude enzyme liquid
70～80 parts in corn stalk, core, 20～30 parts in wheat bran, 1～2 part in ammonium sulfate, profit water, material: water=1: 2.8～3.0, sterilization connects 5～7 parts of aspergillus niger An76 second class inoculums, at 25～28 ℃, aseptic condition bottom fermentation 4～6 days, relative air humidity 80～85%, cryodrying below 80 ℃ is pulverized; Fermenting culture is in 1: the ratio of (4～6) was soaked 10～13 hours, and Plate Filtration, supernatant liquor are the crude enzyme liquid of zytase.More than be weight part, weight ratio.
2. enzymolysis corn stalk (core) prepares wood oligose
(1) processing of raw material
Select for use the corn stalk (core) that is rich in xylan to be raw material, be crushed to 15～20 orders with pulverizer and make particle, particle adds 50% water and mixes thoroughly, the high pressure steam of packing into is sprayed in the quick-fried machine, be forced into 1.2～1.5 MPas, kept 5～15 minutes, reduce to normal barometric pressure suddenly, make the particle explosion become the loose powder shape, prepare the raw material of xylan as enzymolysis with this.
(2) wood oligose preparation
The zytase crude enzyme liquid with through steam explosion pretreated hemicellulose raw material according to (10～11): 1 mixed, 45～50 ℃ of enzymolysis 6～9 hours.Enzymolysis solution is through Plate Filtration, and the supernatant liquor low-voltage vacuum is concentrated into 25～35% of original volume, obtains the wood oligose concentrated solution.
3. absorption
Oligosaccharides concentrated solution and 1: 1 weight ratio of Endomycopsis Fibnligera yeast culture are adsorbed, and adopt the oven dry of low temperature fluidized-bed process, and its gas flow temperature is no more than 90 ℃.
At last, through pulverizing, quality inspection, packing cost invention product.
The bacterial classification of above-mentioned aspergillus niger An-76, Endomycopsis Fibnligera is made can be by prior art.The invention provides following concrete operations step:
A. slant strains (solid)
20～30 parts of potato decortications are cleaned, cut into pieces, be placed in the water potato: water=1: (3～4) weight ratio, 80 ± 1 ℃ were soaked 1 hour, or boiled half an hour, boiled filtration with filter cloth, filtrate thin up to 100 part.Sugaring is 1～2 part again, 1.5～2 parts in agar, natural pH value, 8 pounds of sterilizations.Streak inoculation, 28～30 ℃ of cultivations.
B. second class inoculum (liquid)
The same inclined-plane of culture medium prescription does not only add agar, natural pH value, and bottling, 8 pounds of sterilizations connect bacteria suspension 3～5% weight percents of slant strains, under 28～30 ℃, the shaking table cultivation.
Excellent results of the present invention is: integrate enzyme engineering technology and fermentation engineering, the invention product has the dual-use function of active bacteria formulation and two qi factors.Yeast viable bacteria in the product breeds rapidly at the leading portion of animal digestive tract, can be fast that the oxygen depletion in the digestive tube is intact, in enteron aisle, form anaerobic environment.Wood oligose in the product then, as two qi factors, the propagation of bifidus bacillus in can narrow spectrum promotion enteron aisle, thereby the propagation that suppresses various pathogenic bacterias, improve animal immunologic function, improve the gastral microecological balance of animal, reduce the use of antibiotic medicine, thereby can produce organic animal products of antibiotic-free drug residue.Crop stalk is one of huge agricultural wastes resource of reserves, up to the present, because there is not effective application technology as the second resource, make that the utilization ratio of this resource is very low, annual a large amount of straws are burned, not only waste resource, and air has been caused serious pollution, become the present problem demanding prompt solution of China.This technology in development and use straw resource, no waste discharge, avoid straw burn to environment cause severe contamination, be the effective way of an agricultural wastes resource utilization, high-valued, eco-utilization.
(4) embodiment
Embodiment 1.
The bacterial classification of aspergillus niger An-76, Endomycopsis Fibnligera is made:
A. slant strains (solid)
20 parts of potato decortications are cleaned, cut into pieces, be placed in the water potato: water=1: 4 weight ratio, 80 ± 1 ℃ were soaked 1 hour, boiled filtration with filter cloth, filtrate thin up to 100 part.Sugaring is 2 parts again, 2 parts in agar, natural pH value, 8 pounds of sterilizations.Streak inoculation, 29 ℃ of cultivations.
B. second class inoculum (liquid)
The same A of culture medium prescription does not only add agar, natural pH value, and bottling, 8 pounds of sterilizations connect bacteria suspension 4% weight percent of slant strains, under 29 ℃, the shaking table cultivation.
The Endomycopsis Fibnligera yeast culture is made:
With the Endomycopsis Fibnligera is bacterial strain, and with 35 parts of dregs of beans, 55 parts of corns, 25 parts in wheat bran, ammonium sulfate is substratum for 1.5 parts, raw material mixes through pulverizing, profit water, water content 60%, sterilization, connect 6 parts of Endomycopsis Fibnligera second class inoculums, shallow-layer fermentation 23 hours, 29 ℃ of temperature, relative humidity 82%, 60 ℃ of dryings are pulverized.More than be weight part.
1. the preparation of zytase crude enzyme liquid
75 parts of corn stalk, 28 parts in wheat bran, 1.5 parts in ammonium sulfate, profit water, material: water=1: 3.0, sterilization connects 6 parts of aspergillus niger An76 second class inoculums, in 25 ℃ of aseptic condition bottom fermentations 5 days, 82%, 50 ℃ of drying of relative air humidity, pulverizes; Fermenting culture was soaked 12 hours in 1: 5 ratio, and Plate Filtration, supernatant liquor are the crude enzyme liquid of zytase.More than be weight part, weight ratio.
2. the enzymolysis corn stalk prepares wood oligose
20 order corn stalk powders, particle add 50% water to be mixed thoroughly, and the high pressure steam of packing into is sprayed in the quick-fried machine, is forced into 1.3 MPas, keeps 10 minutes, reduces to normal barometric pressure suddenly, makes the particle explosion become the loose powder shape, prepares the raw material of xylan as enzymolysis.The zytase crude enzyme liquid with through steam explosion pretreated hemicellulose raw material mixed according to 10: 1,48 ℃ of enzymolysis 8 hours.Enzymolysis solution is through Plate Filtration, and the supernatant liquor low-voltage vacuum is concentrated into 30% of original volume, obtains the wood oligose concentrated solution.
3. absorption
Oligosaccharides concentrated solution and 1: 1 weight ratio of Endomycopsis Fibnligera yeast culture are adsorbed, and adopt the oven dry of low temperature fluidized-bed process, 80 ℃ of its gas flow temperatures.
At last, through pulverizing, quality inspection, packing cost invention product.
Embodiment 2. is as described in the embodiment 1, and different is
1. the preparation of zytase crude enzyme liquid
70 parts of corn cobs, 25 parts in wheat bran, 2 parts in ammonium sulfate, profit water, material: water=1: 2.8, sterilization connects 5 parts of aspergillus niger An76 second class inoculums, in 27 ℃ of aseptic condition bottom fermentations 5.5 days, 84%, 60 ℃ of drying of relative air humidity, pulverizes; Fermenting culture was soaked 12 hours in 1: 5 ratio, and Plate Filtration, supernatant liquor are the crude enzyme liquid of zytase.
2. the enzymolysis corn cob prepares wood oligose
With pulverizer corn cob meal is broken to 20 order corn cob meals, particle adds 50% water and mixes thoroughly, the high pressure steam of packing into is sprayed in the quick-fried machine, be forced into 1.5 MPas, kept 12 minutes, and reduced to normal barometric pressure suddenly, make the particle explosion become the loose powder shape, with the zytase crude enzyme liquid in 1 and loose powder shape corn cob particle mixed according to 11: 1,50 ℃ of enzymolysis 8 hours.Enzymolysis solution after filtration, the supernatant liquor low-voltage vacuum is concentrated into 30% of original volume, obtains the wood oligose concentrated solution.
3. absorption
Oligosaccharides concentrated solution and 1: 1 weight ratio of Endomycopsis Fibnligera yeast culture are adsorbed, and adopt the oven dry of low temperature fluidized-bed process, 70 ℃ of its gas flow temperatures.
The outward appearance of the composite microorganism type forage additives that is rich in wood oligose of the embodiment of the invention is a brown, the fermented product special odor is arranged, powdery; This product is rich in wood oligose and yeast cell, wood oligose 〉=10%, and yeast 〉=500,000/g is rich in vitamin B group simultaneously.The present invention is not limited only to the foregoing description.
Claims (3)
1. a preparation method who is rich in the composite microorganism type forage additives of wood oligose is characterized in that, comprises the steps: the preparation of (1) zytase crude enzyme liquid: 70～80 parts in corn stalk, core, 20～30 parts in wheat bran, 1～2 part in ammonium sulfate, profit water, material: water=1: 2.8～3.0, sterilization connects 5～7 parts of aspergillus niger An76 second class inoculums, at 25～28 ℃, aseptic condition bottom fermentation 4～6 days, relative air humidity 80～85%, cryodrying below 80 ℃ is pulverized; Fermenting culture is in 1: the ratio of (4～6) was soaked 10～13 hours, filtered, and supernatant liquor is the crude enzyme liquid of zytase, more than is weight part, weight ratio;
(2) enzymolysis corn stalk, core prepare wood oligose: the zytase crude enzyme liquid with through steam explosion pretreated hemicellulose raw material according to (10～11): 1 mixed, 45～50 ℃ of enzymolysis 6～9 hours, enzymolysis solution after filtration, the supernatant liquor low-voltage vacuum is concentrated into 25～35% of original volume, obtains the wood oligose concentrated solution;
(3) absorption: wood oligose concentrated solution and Endomycopsis Fibnligera yeast culture adsorb by 1: 1 weight ratio, adopt the oven dry of low temperature fluidized-bed process, and its gas flow temperature is no more than 90 ℃.
2. the preparation method who is rich in the composite microorganism type forage additives of wood oligose as claimed in claim 1, it is characterized in that, the pre-treatment of described raw material is, with pulverizer corn stalk, core raw material are crushed to 15～20 orders, particle adds 50% water to be mixed thoroughly, goes into high pressure steam and sprays in the quick-fried machine, be forced into 1.2～1.5 MPas, kept 5～15 minutes, bust makes the particle explosion become the loose powder shape to normal barometric pressure again.
3. the preparation method who is rich in the composite microorganism type forage additives of wood oligose as claimed in claim 1, it is characterized in that, described Endomycopsis Fibnligera yeast culture is to be bacterial strain with the Endomycopsis Fibnligera, with 30～40 parts of dregs of beans, 50～60 parts of corns, 20～30 parts in wheat bran, ammonium sulfate is substratum for 1～2 part, profit water water content 55～65%, sterilization, connect 5～7 parts of Endomycopsis Fibnligera second class inoculums, shallow-layer fermentation 20～24 hours, 28～30 ℃ of temperature, relative humidity 80～85%, cryodrying below 80 ℃, pulverizing and get are weight part.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011274425A CN1147581C (en) | 2001-09-26 | 2001-09-26 | Preparation method of complex microecological feed additive richly containing wood oligose |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CNB011274425A CN1147581C (en) | 2001-09-26 | 2001-09-26 | Preparation method of complex microecological feed additive richly containing wood oligose |
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| CN1341372A CN1341372A (en) | 2002-03-27 |
| CN1147581C true CN1147581C (en) | 2004-04-28 |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100448979C (en) * | 2005-12-09 | 2009-01-07 | 山东省中协食品添加剂研究开发中心 | Production method for xylose by enzyme process |
| CN101912154B (en) * | 2010-08-13 | 2012-07-04 | 川渝中烟工业有限责任公司 | Method for preparing tobacco shred substitute by using tobacco stem surface |
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2001
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