CN104232728B

CN104232728B - Rapeseed peptide (rsp) albumen and preparation method thereof

Info

Publication number: CN104232728B
Application number: CN201410464982.XA
Authority: CN
Inventors: 邓小晨; 吴谦
Original assignee: Sichuan University
Current assignee: Sichuan University
Priority date: 2014-09-12
Filing date: 2014-09-12
Publication date: 2017-07-07
Anticipated expiration: 2034-09-12
Also published as: CN104232728A

Abstract

The invention belongs to technical field of microbial fermentation, and in particular to a kind of rapeseed peptide (rsp) albumen, formed by the fermenting raw materials of following weight proportion：97~99 parts of rapeseed dregs powder, 0~1.5 part of corn flour, 0.4~1.5 part of mixed yeast solid spawn, 0.1~0.4 part of nutritive salt, 0.03~0.1 part of mixing enzyme preparation, 0.1~0.3 part of Bacillus licheniformis liquid, water；Wherein, bacillus licheniformis bacterium solution accounts for other five kinds the 0.6~1.2 of raw material gross weight times with the gross weight of water.The preparation method of rapeseed peptide (rsp) albumen, comprises the following steps：Take each raw material to be well mixed, ferment 18~24h under anaerobic state in 28~32 DEG C, then in the 30~40h that fermented at 28~35 DEG C under aerobic state, dries.The vegetable seed cake protein can be advantageously applied in the raising of domestic birds and animals.

Description

Rapeseed peptide (rsp) albumen and preparation method thereof

Technical field

The invention belongs to technical field of microbial fermentation, and in particular to a kind of rapeseed peptide (rsp) albumen and preparation method thereof.

Background technology

Current China aquaculture heavy dependence corn-soybean meal diet, particularly main protein raw material dregs of beans, portion big absolutely It is divided into import, is limited by international market, often results in aquaculture cost fluctuation, risk increase.In addition, contain in import dregs of beans turning Gene element, does not meet the requirement of organic farm products.

Rapeseed dregs is the accessory substance after rapeseed oil, and according to the difference of process equipment, rapeseed dregs is divided into 95 type vegetable seed again The dregs of rice and 200 type rapeseed dregs.Due to digesting and assimilating with the index such as albumen solubility less than 200 types for 95 type rapeseed dregs, bitter is agreeable to the taste Property it is worse, application in feed is far below 200 type rapeseed dregs.

Rapeseed dregs rich in nutrition content, crude protein content 36% or so, amino acid composition is relatively balanced, the sulphur amino such as methionine Acid content is higher than dregs of beans.But rapeseed dregs contains more noxious material and ANFs, such as：Isothiocyanates, thiocyanates, evil The materials such as zolidine thione, nitrile, sinapine, tannin, phytic acid, not only influence daily ration palatability, influence other nutrient utilizations, also Animal Thyroid Gland Swell can be caused, liver damage suppresses growth of animal, and the death rate rises etc., so as to significantly limit its Application in animal diets.Overall utilization in feed is less than 30%.

Using non-detoxification treatment rapeseed dregs, in animal and fowl fodder is prepared, cub fowl does not eat rapeseed dregs, broiler chicken consumption typically Below 10%, laying hen, breeder are 8% or so；Grower pigs control below 5%, sow below 3%,

In order to remove the poisonous and harmful substance in rapeseed dregs, those skilled in the art once by Physical, chemical method and Bioanalysis is processed, and see the table below 1.

The physics of table 1, chemistry and Biochemical method rapeseed dregs compare

Physics and chemical method can partly remove toxic component, but can not improve nutritive value, can cause on the contrary The loss of nutrient, has abandoned.Most have application value is microbe fermentation method.

The bioanalysis reported in the past uses merely fermentation process, is divided into Unareobic fermentation and the class of aerobic fermentation method two.

The traditional fermentation process of table 2 treatment rapeseed dregs compares

	Unareobic fermentation	Aerobic fermentation method
			Technological process	Anaerobic fermentation	Aerobic fermentation
Total fermentation time	2~4 weeks	≤48h
			Sulphur glucoside is removed	>=80%	>=65%
ANFs is degraded	It is few	It is few
			Crude protein after treatment	36% is constant	~44% improves

Although mentioned microorganism method can remove giucosinolate, trophic factors can not be effective against and dropped Solution, while crude protein is also without too big change, so, searching one kind is badly in need of in this area can degrade anti-battalion in rapeseed dregs simultaneously The method for supporting the factor and giucosinolate, so that for effective utilization of rapeseed dregs provides a better way.

The content of the invention

For the defect that existing microbe fermentation method is present, the present invention provides a kind of rapeseed peptide (rsp) albumen and its preparation side Method.This process employs kluyveromyces marxianus Kluyveromyces marxianus, the bacterial strain is on July 7th, 2014 China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC) is preserved in, deposit number is CGMCC No.9427, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, postcode 10010.Be combined for anaerobic fermentation and aerobic fermentation by the inventive method, while adding appropriate enzyme cooperation treatment, is improving dish Significantly reduce ANFs in the seed dregs of rice while giucosinolate clearance again and improve protein digestibility, so that pole The earth improves the use value of rapeseed dregs.

First technical problem to be solved by this invention is to provide a kind of rapeseed peptide (rsp) albumen.The rapeseed peptide (rsp) albumen is by following The fermenting raw materials of weight proportion are formed：97~99 parts of rapeseed dregs powder, 0~1.5 part of corn flour, mixed yeast solid spawn 0.4 ~1.5 parts, 0.1~0.4 part of nutritive salt, 0.03~0.1 part of mixing enzyme preparation, 0.1~0.3 part of Bacillus licheniformis liquid, water； Wherein, bacillus licheniformis bacterium solution accounts for other five kinds the 0.6~1.2 of raw material gross weight times with the gross weight of water.

Preferably, above-mentioned rapeseed peptide (rsp) albumen is formed by the fermenting raw materials of following weight proportion：98 parts of rapeseed dregs powder, corn flour 0.5 part, 1.2 parts of mixed yeast solid spawn, 0.25 part of nutritive salt, 0.05 part of mixing enzyme preparation, Bacillus licheniformis liquid 0.15 part, water；Wherein, bacillus licheniformis bacterium solution accounts for other five kinds the 0.9 of raw material gross weight times with the gross weight of water.

Preferably, the rapeseed dregs powder described in above-mentioned rapeseed peptide (rsp) albumen is in 95 type rapeseed dregs powders or 200 type rapeseed dregs powders Any one.

It is further preferred that the rapeseed dregs powder described in above-mentioned rapeseed peptide (rsp) albumen is 95 type rapeseed dregs powders.

Preferably, the granularity of the rapeseed dregs powder described in above-mentioned rapeseed peptide (rsp) albumen was 1.8~2.0mm sieve apertures.

Specifically, the preparation method of the bacillus licheniformis bacterium solution described in above-mentioned rapeseed peptide (rsp) albumen is：From lichens gemma In the slant preservation strain of bacillus (Bacillus licheniformis) in picking strain access meat soup protein culture medium, 34~38 DEG C of 30~40h of culture, obtain final product bacillus licheniformis bacterium solution.

Specifically, the preparation method of the mixed yeast solid spawn described in above-mentioned rapeseed peptide (rsp) albumen is：It is false from protein is produced In the slant preservation strain of silk saccharomycete (Candida utilis) in picking strain access potato nutrient solution, in 28~32 DEG C of temperature Degree 2~3d of culture, obtains candida utili bacteria culture fluid；From kluyveromyces marxianus bacterium (Kluyveromyces Marxianus during picking strain accesses potato nutrient solution in slant preservation strain), in 28~32 DEG C of temperature 2~3d of culture, obtain To kluyveromyces marxianus bacteria culture fluid；Again by weight, candida utili Jun Pei Yang Ye ︰ kluyveromyces marxianus bacterium The mixing of nutrient solution=0.75~0.9 ︰ 0.1~0.25 is accessed in potato nutrient solution, in 28~32 DEG C of temperature 2~3d of culture, is obtained final product To compound barm liquid；Compound barm liquid is transferred in wheat bran corn culture medium in 28~32 DEG C of temperature 2~3d of culture again, is obtained final product mixed Close saccharomycete solid spawn.

Preferably, in the preparation method of above-mentioned mixed yeast solid spawn, candida utili bacteria culture fluid and mark This kluyveromyces bacteria culture fluid gross weight is 0.5~1.5 ︰ 100 with potato nutrient solution weight ratio.

Preferably, in the preparation method of above-mentioned mixed yeast solid spawn, by weight, compound barm Ye ︰ wheat brans Corn culture medium=0.5~1.5 ︰ 100.

Preferably, in the preparation method of above-mentioned mixed yeast solid spawn, the candida utili Jun Pei Yang Ye ︰ horses Gram this kluyveromyces bacteria culture fluid=0.85 ︰ 0.15.

Specifically, the preparation method of the wheat bran corn culture medium described in above-mentioned rapeseed peptide (rsp) albumen is：Count by weight ratio, The ︰ 8 of Fu Pi ︰ Yu meter ︰ water=7~8 ︰ 2~3 mix after the 25~40min that sterilized at 118~126 DEG C, cooling three.

Specifically, the nutritive salt described in above-mentioned rapeseed peptide (rsp) albumen includes following component：Meter takes 1 part by weight ratio KH₂PO₄, 0.8 part of KCl, 0.2 part of MgSO₄, 0.05 part of FeSO₄With 0.025 part of ZnSO₄。

Specifically, the mixing enzyme preparation described in above-mentioned rapeseed peptide (rsp) albumen includes following component：Count by weight ratio, phytic acid The ︰ 0.4 of 0.3~0.8 ︰ of Mei ︰ Xian dimension Su Mei ︰ Mu Ju Tang Mei ︰ Guo Jiao Mei ︰ 0.5~1.5 ︰ of papain=8~12 ︰ 0.8~1.5 ~0.6.

Preferably, the mixing enzyme preparation described in above-mentioned rapeseed peptide (rsp) albumen is meter Zhi Suan Mei ︰ Xian dimension Su Mei ︰ by weight ratio The ︰ 0.5 of 0.5 ︰ of Mu Ju Tang Mei ︰ Guo Jiao Mei ︰ 1 ︰ of papain=10 ︰ 1.

Specifically, the enzyme activity of phytase described in above-mentioned rapeseed peptide (rsp) albumen is 5000 units/g.The cellulase Enzyme activity is 1000 units/g.The enzyme activity of the zytase is 100,000 units/g.The enzyme activity of the pectase is 10,000 single Position/g.The enzyme activity of the papain is 800,000 units/g.

Further, kluyveromyces marxianus Kluyveromyces marxianus in above-mentioned rapeseed peptide (rsp) albumen Preserving number is the bacterial strain of CGMCC No.9427.

Specifically, the preparation method of above-mentioned rapeseed peptide (rsp) albumen is：

A, mixing：Take rapeseed dregs powder, corn flour, Bacillus licheniformis liquid, mixed yeast solid spawn, nutritive salt, Mixing enzyme preparation and water mix to obtain fermentation material；

B, fermentation：After fermentation material is fermented into 18~24h in 28~32 DEG C under anaerobic state, take out, then in aerobic state Under at 28~35 DEG C ferment 30~40h, dry after i.e. obtain rapeseed peptide (rsp) albumen.

Second technical problem to be solved by this invention is to provide a kind of preparation method of above-mentioned rapeseed peptide (rsp) albumen.The party Method is comprised the following steps：

Preferably, in above-mentioned preparation method step B, the thickness of fermentation material is 400~500mm when anaerobic state ferments.

Preferably, in above-mentioned preparation method step B, the container that anaerobic fermentation is used is stainless steel.

It is further preferred that in above-mentioned preparation method step B, the container that anaerobic fermentation is used for length, width and height 2000mm × The stainless steel metal case of 1000mm × 500mm.

Preferably, in above-mentioned preparation method step B, the thickness of fermentation material is no more than 60mm during aerobic state fermentations.

Preferably, in above-mentioned preparation method step B, the container that aerobic fermentation is used is appointed for stainless steel or bamboo Meaning is a kind of.

It is further preferred that in above-mentioned preparation method step B, the container that aerobic fermentation is used for length, width and height 2000mm × The stainless steel metal tray of 1000mm × 80mm.

Further, the preserving number of the kluyveromyces marxianus Kluyveromyces marxianus in the above method It is the bacterial strain of CGMCC No.9427.

Compared with untreated rapeseed dregs (crude protein 36%), under identical Water Content Conditions, crude protein can for product of the present invention Up to 43~44%, total amino acid content improves 16%, giucosinolate clearance 75~80%, phytic acid degradation rate about 45%, fruit The ANFs such as glue degradation rate 40%, fiber and tannin are also reduced in various degree, and acid-soluble protein content increases by 100%, greatly Width improves Feeding Value.Aquaculture cost, additional income can be reduced using product of the present invention；Can be improved using product of the present invention Animal body and intestinal health level, strengthen premunition, routinely without antibiotic can be conducive to carrying out nonreactive healthy aquaculture； By ferment treatment and fermentation, rich in growth factor and peptides etc., the qualities such as meat egg can be improved, improve agricultural product quality；Improve Breeding environment, is conducive to decontamination of human excreta and resource, and development cleaning is cultivated and ecological circulation.

Kluyveromyces marxianus Kluyveromyces marxianus are make use of in the present invention, the bacterial strain is in 2014 On July 7, in is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), and deposit number is CGMCC No.9427, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, postal Compile 10010.

Specific embodiment

Rapeseed dregs due to containing poisonous and harmful substance (such as giucosinolate), ANFs (such as fiber, phytic acid, pectin, Tannin etc.), it is impossible to it is directly used in the edible of domestic birds and animals, it is necessary to detoxification is carried out to rapeseed dregs and ANFs treatment is eliminated.

Prior art wants simultaneously simultaneous mainly using Physical, chemical method and microbe fermentation method treatment rapeseed dregs Turn round and look at removing rapeseed dregs in giucosinolate and eliminate ANFs so that improve digestibility, improve nutritive value and Feeding effect, prior art is extremely difficult to above-mentioned desired effect.To solve above-mentioned difficulties, prior art is mainly using complexity Bacterial strain is combined, and adds bacterial strain to decompose corresponding material, but these methods often run counter to Microbiological Principle, and bacterial strain is too much often Contaminated bacteria each other, influences each other, potential difference long；Or these bacterial strains optimum growing condition difference in itself is larger, in same matrix Middle growth is interfered.So in order to be able under same matrix condition to the giucosinolate in rapeseed dregs, phytic acid, fibre object Matter, pectin and tannin etc. can well carry out resolution process, it is necessary to reasonably select bacterial strain, could be to the sulphur in rapeseed dregs Glycoside, phytic acid, fibrous matter, pectin and tannin etc. carry out good working process.

So, due to nature can degrade giucosinolate bacterial strain it is more, it is necessary to by many experiments screening could obtain To the strain excellent for processing giucosinolate in rapeseed dregs.The present inventor is with bacterium, saccharomycete and filamentous fungi The hundreds of plants of bacterium of three major types microorganism, carry out aerobic culture in rapeseed dregs culture medium, determine increment, the sulphur grape of bacterial strain Glucoside resolution ratio, crude protein content and outward appearance smell, have discharged filamentous fungi；Further, and it is mixed according to bacterial strain and is surveyed Determine the indexs such as increment, giucosinolate resolution ratio, crude protein content and the outward appearance smell of bacterial strain, then consider bacterial strain in itself Security, the complexity of industrialized production, it is final to determine selection candida utili bacterium, bacillus licheniformis and Marx Three kinds of bacterial strains of Kluyveromyces sp carry out processing rapeseed dregs.

It is found through experiments that, in above-mentioned three kinds of bacterial strains, the biomass of candida utili bacterium is high detoxification ability；Mark This Kluyveromyces aroma-producing yeasts, there is faint detoxification ability, but can produce ester perfume, improves fermentation rapeseed dregs palatability；Lichens Bacillus can improve body nospecific immunity and intestinal health, and bacillus licheniformis can be by enteron aisle, and part can enter excrement Just, cleaning is conducive to cultivate；Find simultaneously, kluyveromyces marxianus bacterium and bacillus licheniformis inoculum concentration are crossed conference reduction and produced The detoxification ability of protein candidiasis, accordingly, it would be desirable to the inoculative proportion of these three bacterial strains is accurately determined under industrial process conditions, By substantial amounts of experimental studies have found that, bacillus licheniformis Amplification Culture ties up candida utili bacterium and Marx's Crewe Obtain mixed yeast solid spawn after saccharomycete mixing Amplification Culture, control addition mixed yeast solid spawn for 0.4~ 1.5 parts, Bacillus licheniformis liquid be 0.1~0.3 part when can well decompose sulfur sugar in 97~99 parts of rapeseed dregs powder Glucoside.Preferably 1.2 parts of mixed yeast solid spawn, 0.15 part of Bacillus licheniformis liquid.

It is further discovered that, although above-mentioned three kinds of bacterial strain combined fermentations can decompose giucosinolate, but to anti-nutrition of degrading The factor and the raising effect of digesting and assimilating are limited.The presence of ANFs has a strong impact on the application of rapeseed dregs, for example：Phytic acid is dish Main ANFs in the seed dregs of rice, about 60~70% phosphorus is combined in phytic acid in rapeseed dregs, and hardly possible is ruminated by non-very much Livestock and poultry animal utilize.Because the phosphorus in phytic acid is not absorbed, discharged by enteron aisle, so as to improve phosphorus content in excrement, caused The possibility of phosphorus contaminant water increases.Also, due to the strong chelation of phytic acid, can securely chelate positively charged Zinc, Iron, Calcium copper The divalence such as magnesium manganese or polyvalent metal ion, form insoluble phytates chelate, so that the life of some essential mineral elements Thing potency is substantially reduced.Therefore, in rapeseed dregs phytic acid degraded, be also rapeseed dregs treatment emphasis.

In order to solve the problem of degraded ANFs, the present inventor is by flat board transparent circle method in substantial amounts of bacterium The phytase producing strains and a kind of phytase producing strains of bacterium class of two primary yeast classes have been selected in kind.In order to same Giucosinolate and ANFs are processed simultaneously in matrix, the present inventor is also contemplated between various strains Be mutually combined in the case of to giucosinolate and the degraded situation of phytic acid.It has been investigated that：With the increasing of bacterial strain number of combinations Plus, disturb obvious between bacterial strain, so as to the degradation rate for causing phytic acid and giucosinolate largely declines.Also, in Mixed Microbes Under the fermentation condition of strain fermentation and pure rapeseed dregs, nor the optimum condition of phytase producing strains phytase generating, so as to cause to plant Sour production of enzyme is low, and P elements burst size is few.Also, production of enzyme Pectinase Producing Strain higher and cellulase producing bacteria are usually Filamentous fungi, producing enzyme is also very low in rapeseed dregs base-material, also with mould taste.

In face of the problem of the above-mentioned ANFs that needs to degrade, inventor is again by be further discovered that after many experiments can be with Add complex enzyme formulation to solve the above problems, be found through experiments that using cellulase, phytase, pectase and zytase Collective effect can not only decompose Main Antinutritional Factors in rapeseed dregs, moreover it is possible to improve the detoxification ability of microorganism；Further send out Existing, enzyme compound use has certain synergy, such as phytic acid content is higher in rapeseed dregs, decomposition of the phytase to phytic acid, can Combination of the phytic acid to nutrient is reduced, the phosphorus of release is decomposed again for growth of microorganism provides nutrient from phytic acid, be conducive to micro- life Thing grows, and the catabolite of pectin and xylan etc. can also provide the nutrient of growth of microorganism in addition.

The present inventor is investigated influence of the addition of phytase to bacterial strain removing giucosinolate, as a result sends out It is existing, when the amount of phytase is controlled in OK range, can well remove giucosinolate and degraded phytic acid；While inventor It is investigated influencing each other for various enzyme preparations.By considering after various researchs when mixing enzyme preparation includes following composition When Zhi Suan Mei ︰ Xian dimension Su Mei ︰ Mu Ju Tang Mei ︰ Guo Jiao Mei ︰ papain=8~12 ︰ 0.5~1.5 ︰ 0.3~0.8 ︰ 0.8~ 1.5 ︰ 0.4~0.6, not only can well process the ANFs in rapeseed dregs, additionally it is possible to make the bacterial strain of addition to sulphur Portugal The degradation rate of grape glucoside is big.It is preferred that the ︰ of 0.5 ︰ of Zhi Suan Mei ︰ Xian dimension Su Mei ︰ Mu Ju Tang Mei ︰ Guo Jiao Mei ︰ 1 ︰ of papain=10 ︰ 1 0.5, as a result find, the sour molten albumen of rapeseed peptide (rsp) albumen improves about 100% than original rapeseed dregs, wherein about 80% is by digesting rank Duan Gongxian's, to giucosinolate virus elimination rate from original simple fermentation when 50~60%, bring up to 70~80%.Wherein, institute The phytase for using is 5000 units for every gram of enzyme activity of enzyme.The cellulase for being used is single for every gram of enzyme activity of enzyme 1000 Position.The zytase for being used is every gram of unit of the enzyme activity of enzyme 100,000.The pectase for being used is every gram of enzyme activity of enzyme 10,000 Unit.The papain for being used is 800,000 units for every gram of enzyme activity of enzyme.

In order to reach good degradation effect, not only to consider the selection of various raw materials, the addition of each raw material, The matching relationship of each raw material, it is necessary to make each compound play preferably effect in specified conditions, so, in order that the bacterium of addition The effect that strain, mixing enzyme preparation are bringd into play, one suitable condition of selection is also essential.

The aerobic fermentation time is short in the prior art, biomass is high, in fermentation process dehydration it is very fast, so as to be unfavorable for that enzyme is made With, therefore be only difficult raising by aerobic fermentation and digest and assimilate.The anaerobic fermentation time is very long, and crude protein and amino acid do not increase, raw Performance long is general, and water content is high after fermentation, it is more difficult to store, and is only suitable for that plant is fresh to feed, and is not suitable for feed manufacturing.So, in order to The bacterial strain and enzyme preparation of addition can be made to be played a role well in same system, degraded giucosinolate and anti-nutrition because Son, it is necessary to which finding rational fermentation process just can be with solve problem.Found by many experiments, processing mode is from simple microorganism Fermentation is changed into the mode that ferment treatment is combined with microbial fermentation, and processing with enzyme preparation does not need oxygen, so as to control previous stage It is anaerobic fermentation, the latter half is aerobic fermentation.In anaerobic fermentation stage, the compound action of various enzymes makes rapeseed dregs knot of tissue Structure can be disintegrated to a certain degree is degraded with corresponding material, so that the ANFs in rapeseed dregs is reduced, is improve The molten protein content of acid, also increases the nutrient for promoting growth of microorganism in rapeseed dregs raw material, such as P elements, de- beneficial to the later stage Poison and raising digestibility；In the aerobic fermentation stage, cell is largely bred under aerobic conditions, so as to promote toxicity sulphur grape Glucoside reduction.

The preparation method of above-mentioned rapeseed peptide (rsp) albumen, comprises the following steps：

Take 97~99 parts of rapeseed dregs powder, 0~1.5 part of corn flour, 0.4~1.5 part of mixed yeast solid spawn, nutritive salt 0.1~0.4 part, 0.03~0.1 part of mixing enzyme preparation, 0.1~0.3 part of Bacillus licheniformis liquid, water；Wherein, lichens brood cell Bacillus bacterium solution accounts for other five kinds the 0.6~1.2 of raw material gross weight times with the gross weight of water；Must be fermented after above-mentioned each material is mixed Material；After fermentation material is fermented into 18~24h in 28~32 DEG C under anaerobic state, take out, then in 28~35 DEG C under aerobic state 30~40h of lower fermentation, rapeseed peptide (rsp) albumen is obtained after drying.

Inventor has found：While anaerobic stages add enzyme preparation, strain inoculum concentration should be controlled, enzymolysis process is also Strain amplification procedure, bacterium is increased by early stage in advance, and strain cell reaches finite concentration, and into after aerobic fermentation, bacterial strain enters immediately Enter fast breeding, in anaerobic stages, inoculation amount can not be excessive, in order to avoid material temperature is raised, and consume too many nutrient, be unfavorable for The biomass accumulation of aerobe fermentation, inoculum concentration very little, can extend total fermentation time.

Preferably, the preparation method of the bacillus licheniformis bacterium solution is：From the slant preservation strain of bacillus licheniformis Middle picking strain is accessed in meat soup protein culture medium, and 30~40h is cultivated at 34~38 DEG C, obtains final product bacillus licheniformis bacterium solution. When bacterial strain, strain has reached a growth stationary phase in the range of this temperature and time, so, can be without considered critical bacterial strain The amount of inoculum concentration and culture medium, similarly hereinafter.

Heretofore described meat soup peptone is the culture medium that microorganism field is commonly used, and preparation method is：Peptone 10g, beef extract 6g, NaCl5g, water 1000mL, after all the components dissolving, the pH value for adjusting the solution with 2NNaOH solution is 7.2 ~7.4, sterilize 30min at a temperature of 121 DEG C, cools down standby.

Preferably, the preparation method of the mixed yeast solid spawn is：Respectively from candida utili bacterium and mark Each picking strain, is respectively connected in potato nutrient solution in the slant preservation strain of this Kluyveromyces sp, in 28~32 DEG C of temperature 2~3d of culture, respectively obtains candida utili bacteria culture fluid and kluyveromyces marxianus bacteria culture fluid；Again by weight, Candida utili Jun Pei Yang Ye ︰ kluyveromyces marxianus bacteria culture fluid=0.75~0.9 ︰ 0.1~0.25 is (preferably 0.85 ︰ 0.15) mixing access potato nutrient solution in, in 28~32 DEG C of temperature 2~3d of culture, that is, obtain compound barm liquid；Wherein, Candida utili bacteria culture fluid and kluyveromyces marxianus bacteria culture fluid gross weight and potato nutrient solution weight ratio be 0.5~ 1.5 ︰ 100 (preferably 1 ︰ 100)；Again by weight, compound barm Ye ︰ wheat brans corn culture medium=0.5~1.5 ︰ 100 is (excellent Elect 1 ︰ 100 as) compound barm liquid is transferred in wheat bran corn culture medium in 28~32 DEG C of temperature 2~3d of culture, obtain final product mixing ferment Female bacterium solid spawn.

Heretofore described potato nutrient solution is that microorganism field often uses culture medium, and preparation method is：By fresh potato It is cut into small pieces after peeling, adds the water of 6 times of potato block gross weight, boil 20min, filter off solid slag, takes supernatant and add The sucrose of amount of liquid 2%, sterilize 30min at a temperature of 121 DEG C, cools down standby.

Water is added to be because while not needing oxygen in anaerobic stages enzymolysis described in raw material of the present invention, but enzymolysis needs Material contains certain moisture, is studied through inventor and found, the water of addition and bacillus licheniformis bacterium solution summation for other five The need for can well meeting enzymolysis to water during 0.6~1.2 times of kind of raw material, while will not be produced to the aerobic fermentation in later stage again Very big influence.

Preferably, early stage is mainly the enzymolysis and addition strain initial stage propagation of mixing enzyme preparation, because enzymolysis does not need oxygen Gas, strain initial stage propagation needs a small amount of oxygen, so it is anaerobic fermentation to control prior fermentation；Meanwhile, enzymolysis and increasing bacterium are required for Water conservation, the too thin easy dehydration of fermentation material, and the container for needing is also more, it is right if too thick can make container too heavy, it has not been convenient to carry Enzymolysis and increasing bacterium influence are little, and the thickness of fermentation material is 400~500mm when being fermented it is advantageous to anaerobic state.

Preferably, the container that anaerobic fermentation is used is stainless steel.

It is further preferred that the container that anaerobic fermentation is used is the stainless steel of length, width and height 2000mm × 1000mm × 500mm Metal box.

Preferably, the later stage be mainly thin layer solid state fermentation, be conducive to improve biomass, be controlled to aerobic fermentation, fermentation material Can not be too thick, it is no more than 60mm it is advantageous to the thickness for fermentation material.

Preferably, the container that aerobic fermentation is used for stainless steel or bamboo any one.

It is further preferred that the preparation method of above-mentioned rapeseed peptide (rsp) albumen, the container that aerobic fermentation is used in step B is length and width The stainless steel metal tray of 2000mm × 1000mm high × 80mm.

The fermentation process of the present invention of table 3 compares with traditional fermentation process treatment rapeseed dregs

In following test example and embodiment, the candida utili for being used comes from Institute of Microorganism, Academia Sinica (Beijing), China General Microbiological culture presevation administrative center (CGNCC), bacterium numbering CGNCC2.281；Bacillus licheniformis From Chinese food fermentation research institute (Beijing), Chinese industrial Microbiological Culture Collection administrative center (CICC), bacterium numbering CICC10098。

Phytase is purchased from Beijing Smistyle Science ＆ Technology Development Co., Ltd., cellulase, pectase, zytase and pawpaw egg White enzyme is purchased from ROTA Bioengineering Co., Ltd..

The screening process of the degraded giucosinolate bacterial strain of test example 1

From nature various raw materials in materials and preservation of bacteria strain, enriched culture separates and has screened former in rapeseed dregs Hundreds of plants of microbial strains of well-grown on material.Through Preliminary Identification, including 15 plants of saccharomycete (numbering Y1~Y15), filamentous fungi 48 plants (numbering M1~M48) and 66 plants of bacterium (numbering B1~B66), see the table below 4：

The flat board primary dcreening operation result of table 4

Numbering

Circle/footpath

Numbering

Circle/footpath

Numbering

Circle/footpath

Numbering

Circle/footpath

Numbering

Circle/footpath

Y1

-

M12

+

M38

+

B16

++

B42

+

Y2

+

M13

-

M39

++

B17

+

B43

-

Y3

+++

M14

+

M40

-

B18

-

B44

-

Y4

-

M15

-

M41

-

B19

++

B45

+

Y5

-

M16

++

M42

+++

B20

+

B46

+++

Y6

+

M17

+

M43

+

B21

-

B47

++

Y7

+++

M18

-

M44

-

B22

+

B48

-

Y8

+

M19

-

M45

++

B23

+++

B49

+

Y9

+

M20

-

M46

+

B24

+

B50

++

Y10

+++

M21

++

M47

+++

B25

++

B51

+

Y11

-

M22

+++

M48

+++

B26

-

B52

++

Y12

++

M23

-

B1

++

B27

-

B53

+

Y13

+++

M24

+

B2

-

B28

++

B54

+++

Y14

+++

M25

+

B3

-

B29

+

B55

++

Y15

+

M26

++

B4

+

B30

+++

B56

-

M1

-

M27

+

B5

++

B31

++

B57

+

M2

-

M28

+++

B6

+

B32

-

B58

-

M3

+

M29

-

B7

-

B33

++

B59

-

M4

-

M30

-

B8

+

B34

+

B60

+

M5

+

M31

++

B9

+

B35

-

B61

-

M6

++

M32

+

B10

++

B36

++

B62

+++

M7

-

M33

+++

B11

+

B37

-

B63

+

M8

++

M34

++

B12

-

B38

-

B64

+

M9

-

M35

+

B13

+

B39

+

B65

-

M10

++

M36

-

B14

++

B40

-

B66

++

M11

+++

M37

+

B15

+

B41

+

Note：In table+~++++, growth good job situation is represented respectively.

As shown in upper table 4, Y3, Y7, Y10, Y13, Y14, M11, M22, M28, M33, M42, M47, M48, B23, B30, This 17 plants of bacteria growing situations of B46, B54, B62 are preferable.

Shaking flask secondary screening experiment is further carried out with rapeseed dregs nutrient solution to the above-mentioned 17 plants of bacterium for filtering out, main result sees below Table 5：

The shaking flask secondary screening of table 5 determines OZT, ITC, GS degradation rate

Numbering	OZT degradation rates (%)	ITC degradation rates (%)	GS degradation rates (%)
				Y3	45.52	46.26	48.92
Y7	66.82	67.41	68.23
				Y10	64.28	67.26	69.15
Y13	50.84	53.97	54.38
				Y14	57.12	58.95	59.46
M11	42.64	42.18	43.66
				M22	38.23	32.01	36.52
M28	53.68	54.23	54.97
				M33	33.54	35.27	39.36
M42	61.23	60.24	63.54
				M47	43.48	42.67	44.39
M48	42.67	43.85	44.61
				B23	48.65	47.25	49.67
B30	42.53	42.76	43.92
				B46	36.39	37.84	39.24
B54	45.32	49.21	57.24
				B62	63.26	64.38	64.45

GS, OZT, ITC in above-mentioned table 5 represent giucosinolate, sulphur glucoside Jiang solution Chan Wu oxazolidines thioketones, different sulphur cyanogen respectively Acid esters；It can be seen that this 10 plants of bacterium of Y3, Y7, Y10, Y13, Y14, M28, M42, B23, B54, B62 are with preferable from above-mentioned table 5 Giucosinolate removal ability.Through detection in addition, wherein M28 has tannin enzymatic activity, though B46 degrades slightly to giucosinolate Difference, but have phytase activity.

In above experimental strain, after testing, Y7 bacterial strains are candida utili bacterium (Candida utilis)；Y13 bacterium Strain is kluyveromyces marxianus bacterium (Kluyveromyces marxianus) through DNA sequencing, and the bacterial strain deposit number is CGMCC No.9427, belong to aroma-producing yeasts；M42 bacterial strains are aspergillus niger, with pectin ability；B6 is bacillus licheniformis, tool There is starch to decompose and protein breakdown ability, virus elimination rate is relatively low.Several bacterium are the safe bacterial strain of feed and food above.Separately Outward, individual plant solid state fermentation is carried out in rapeseed dregs matrix, bacterium B23, B54, B62 smell is more smelly, abandons；Filamentous fungi M28 bands Musty, abandons.

Based on the above results, Y3, Y7, Y13, M42, phytase bacterial strain B46 and bacillus licheniformis B6 is selected to be combined reality Test：

(1) main result of combination of two strain fermentation：

Y3 bacterial strains, Y7 bacterial strains are combined with B6 respectively, when B6 inoculative proportions are (exceeding total inoculum concentration 25%) higher, sulfur sugar Glucoside degradation rate has declined；

Y3 bacterial strains, Y7 bacterial strains are combined with B46 respectively, and phytase generating is very low, and giucosinolate resolution ratio is not apparent from change, but There is weaker commensalism to act between combination bacterial strain, i.e. decomposition of the phytase bacterium producing multi enzyme preparation to phytase is produced P element and can be promoted Enter cell growth；

Y3 bacterial strains, Y7 bacterial strains are combined with M42 respectively, and giucosinolate degradation rate declines as M42 inoculative proportions are improved, Produce pectase extremely low；

Y3 bacterial strains are combined with Y7 bacterial strains, and giucosinolate degradation rate is not lifted, therefore abandon Y3, the known bacterial strain Y7 of choosing；

(2) three plants of bacterial strains and the main result of combination of the above fermentation：

In Y7-B46-B6 combinations, Y7-Y13-B6 combinations, Y7-B6-M42 combinations, Y7-B46-B6-M42 combinations, Y7-Y13- In the combination such as B46-B6-M42, Y7-Y13-B6 combinations relatively preferably, but B46 to the decomposition of phytic acid not as the combination of two bacterial strains, M42 Produce pectase also unobvious.Giucosinolate degraded all declines in four combinations and five combinations, shows, bacterium producing multi enzyme preparation is in rapeseed dregs In matrix and under hybrid bacterial strain fermentation condition, yield of enzyme is extremely low, disturbs bright with the increase of bacterial strain number of combinations, between bacterial strain It is aobvious.

Experimental result is removed according to above giucosinolate, Y7 candida utili and Y13 kluyveromyces marxianus are selected Bacterium (deposit number be CGMCC No.9427), along with B6 bacillus licheniformis totally 3 plants of bacterium.Candida utili Main Function It is detoxification；Kluyveromyces marxianus bacterium Main Function is raw fragrant；Bacillus licheniformis only has faint detoxification efficiency, Main Function It is to improve animal intestinal tract health as probiotics.

In order to avoid being interfered in growth course, it is necessary to accurately control the inoculative proportion of these three bacterial strains.Due to mark This Kluyveromyces sp detoxification efficiency is weaker, and ratio increase can reduce detoxification efficiency, but kluyveromyces marxianus bacterium is raw fragrant Yeast, uses in right amount, can improve fermentate palatability, through experiment, when candida utili bacterium and in kluyveromyces marxianus When bacterium reaches growth balance period in potato nutrient solution respectively, the ratio of each nutrient solution of acquisition is 9~3 ︰ 1, and detoxification efficiency is most It is good, while can also improve the palatability of fermentate, see the table below 6.

Desulfurization glycoside of the ratio of the candida utili of table 6 and kluyveromyces marxianus bacterium to vegetable seed Hectometer raw materials Effect

	1	2	3	4	5	6	7
								Candida utili	10	9	8	7	6	5	4
Kluyveromyces marxianus	—	1	2	3	4	5	6
								De- GS%	53.6	53.4	55.3	54.1	50.0	44.7	41.3

Screening by this test example to bacterial strain, it is final to determine selection candida utili bacterium, kluyveromyces marxianus Bacterium (deposit number is CGMCC No.9427), three kinds of bacterial strains of bacillus licheniformis are come the giucosinolate in rapeseed dregs of degrading.

The screening process of the mixing enzyme preparation of test example 2

Phytic acid is important ANFs in rapeseed dregs, and about 60%~70% phosphorus is combined and planted in rapeseed dregs It is difficult to be utilized by the non-livestock and poultry animal ruminated very much in acid, therefore, phytic acid degraded is only second to giucosinolate, is rapeseed dregs treatment Important content.Because phytate phosphorus are not absorbed, discharged by enteron aisle, so as to improve phosphorus content in excrement, cause phosphorus contaminant water Possibility increases.Further, since the strong chelation of phytic acid, can securely chelate the divalence such as positively charged Zinc, Iron, Calcium copper magnesium manganese Or polyvalent metal ion, insoluble phytates chelate is formed, so that the biological value of some essential mineral elements is obvious Reduce.

By test example 1 it is recognized that while bacterial strain B46 is phytic acid grows bacterium, but in hybrid bacterial strain fermentation and pure rapeseed dregs It is not the optimum condition of bacterial strain B46 phytase generatings under the conditions of base-material top fermentation, phytic acid production of enzyme is very low, and P element burst size is few, Combined by phytase-producing strain and cultivated, increase phytic acid enzyme amount more difficult, it is impossible to it was observed that phytase activity higher is to sulphur grape The influence of glucoside decomposition.In addition, production of enzyme Pectinase Producing Strain higher, tannase and cellulase producing bacteria are usually Filamentous fungi, producing enzyme is also very low in rapeseed dregs base-material, also with mould taste.As combination bacterial strain increases, each other each other Contaminated bacteria, it is difficult to take into account should producing enzyme, the purpose of degrade again various toxicants and ANFs, effect is poor.So, The present invention is by having abandoned the bacterial strains such as selection phytase producing strains come the ANFs in rapeseed peptide (rsp) of degrading after experiment.

Inventor is again by after multi-party thinking and experiment, discovery can add enzyme preparation to reach degraded ANFs Effect.Inventor is in countless enzyme preparation, it is contemplated that the decomposition to various ANFs, and selection adds phytase to be made With.

Inventor considers and demonstrates addition phytase and removes strain growth and sulfur sugar to giucosinolate first The influence of glucoside removal, candida utili is seeded in rapeseed dregs base-material, and adds phytase, as a result see the table below 6：

The influence that the phytase of table 7 is removed and grown to giucosinolate

	Experimental group 1	Experimental group 2	Control group
				GS decomposers	+	+	+
Plus phytase	——	+	——
				Plus KCl	—	+	—
Plus KH₂PO₄	+	—	—
				P incrementss mg/kg*	473	1043	—
GS removes %	55	67	40
				Cell count × 10⁸Individual/ml	5.5	12	3.5

* P incrementss mg/kg, and compares.

From upper table 7, when enough additions phytase (>=0.04%), the increase of P element can be remarkably promoted in rapeseed dregs Cell growth and giucosinolate are removed；Plus KCl is the feedback inhibition for avoiding P element to phytase.The effect of experimental group 2 is best, P element is not added in its compost, decomposition of the P element needed for cell growth from phytase to phytic acid, both reduced anti-nutrition because Son, promotes cell growth, preferable cell growth to increase giucosinolate removal again.

Because the ANFs species in rapeseed dregs is more, after inventor is it is further contemplated that add cellulase, zytase The influence of strain growth and giucosinolate removal is removed to giucosinolate, 8 are the results are shown in Table：

Table 8 is under the conditions of added with phytase, the effect of different cellulase and xylanase content

Experimental group	1	2	3	4	5	6	7	8	9
										Cellulase %	——	0.001%	0.002%	0.002%	0.004%	0.004%	0.008%	0.008%	0.008%
Zytase %	——	0.001%	0.001%	0.002%	0.001%	0.002%	0.002%	0.004%	0.008%
										GS degradation rates	65.34%	62.45%	64.31%	64.54%	72.34%	72.86%	71.23%	73.23%	74.2%

Note：Each group is added with phytase 0.04% (pressing rapeseed dregs base-material)

From upper table 8, experimental group 5 to experimental group 9, giucosinolate degradation rate is higher, but in view of enzyme preparation cost And effect, optimization experiment group 6, i.e. cellulase and zytase addition are respectively 0.004% and 0.002%.

Further, inventor is it is contemplated that in the presence of phytase, cellulase, zytase are common, plus Enter the influence that pectase removes strain growth and giucosinolate removal to giucosinolate, the results are shown in Table 9：

Table 9 is under the conditions of added with phytase, cellulase and zytase, the effect of different pectase contents

Experimental group	1	2	3	4	5	6
							Pectase %	——	0.001%	0.002%	0.005%	0.01%	0.015%
GS degradation rates	71.95%	71.85%	72.44%	76.55%	76.66%	74.23%

Note：Each group (presses rapeseed dregs base added with phytase 0.04%, cellulase 0.004% and zytase 0.002% Material)

From upper table 9, experimental group 4 is high to the degradation rate of giucosinolate when being pectase 0.005%, while pectase Usage amount it is less.

To sum up each table result, determines that the ratio between various enzymes is：Phytase 0.04%, cellulase 0.004%, wood are poly- Carbohydrase 0.002% and pectase 0.04%, the i.e. ︰ 1 of 10 ︰, 1 ︰ 0.5.

On the other hand, because protease can improve rapeseed dregs protein breakdown utilization rate, but protease also can be to other enzyme eggs Its enzyme activity is degraded and reduced in vain.The substrate specificity of different protease is different, and has certain relation with the kind of enzyme source.Through reality Test result to show, separate sources protease has a different to the influence of different hydrolases, general trend be with extended durations of action, Enzymatic activity reduction；In effect 4h, can generally retain more than 50% activity.

From enzyme be easy to get and the mechanism of action etc. considers, selection food-grade papain as subjects, this enzyme Substrate it is specific less strict, under acid, neutral, alkaline environment can decomposing protein, heat resistance is strong.Inventor distinguishes Determine papain to phytase, cellulase, zytase, pectase influence, be as a result displayed in papain In the presence of, these four enzymes can retain more than 50% activity in 4~6h；If by the reduction of Papain enzyme dosage, enzyme activity retention time Can extend.Through experiment, Papain enzyme dosage is about other four kinds 4% or so of enzyme dosage summation, enzyme activity be retained in 50% with About 10h of upper time, the acid-soluble ratio of rapeseed peptide (rsp) albumen can improve 5~10%.So, consider, by weight, final control is planted The ︰ of 0.3~0.8 ︰ of Suan Mei ︰ Xian dimension Su Mei ︰ Mu Ju Tang Mei ︰ Guo Jiao Mei ︰ 0.5~1.5 ︰ of papain=8~12 ︰ 0.8~1.5 0.4~0.6.

In sum, the present invention selects candida utili, (deposit number is CGMCC to kluyveromyces marxianus No.9427), three kinds of bacterial strains of bacillus licheniformis and phytase, cellulase, zytase, pectase, papain five Kind of enzyme preparation come rapeseed dregs of degrading, by ANFs and sulphur grape in the influencing each other of bacterial strain and enzyme preparation, rapeseed dregs The each side such as the condition required for glucoside degradation rate, enzyme preparation effect consider, and it is rapeseed dregs powder 97~99 to control each parameter Part, 0~1.5 part of corn flour, 0.4~1.5 part of mixed yeast solid spawn, 0.1~0.4 part of nutritive salt, mixing enzyme preparation 0.03~0.1 part, 0.1~0.3 part of Bacillus licheniformis liquid, water；Wherein, bacillus licheniformis bacterium solution is accounted for the gross weight of water Other five kinds the 0.6~1.2 of raw material gross weight times.Further, preparation method is in anaerobic state after above-mentioned raw materials are mixed Under in 28~32 DEG C ferment 18~24h after, take out, then under aerobic state at 28~35 DEG C ferment 30~40h, dry after i.e. Obtain rapeseed peptide (rsp) albumen.

Nursing experiment of the preparation and product of the rapeseed peptide (rsp) albumen of embodiment 1 to pig

A, prepare bacillus licheniformis bacterium solution：In strain (CICC10098, similarly hereinafter) being preserved from bacillus licheniformis inclined-plane, Accessed in the triangular flask of 100mL meat soup peptone nutrient solutions with the ring of oese picking 1,35 DEG C culture 2d, then take 20mL transfer into In 2L meat soup peptone nutrient solutions, 2d is cultivated in 35 DEG C, be bacillus licheniformis bacterium solution；

B, prepare mixed yeast solid spawn：From candida utili (CGNCC2.281, similarly hereinafter) and Marx's Crewe Dimension yeast (CGMCC No.9427, similarly hereinafter) inclined-plane is preserved in strain, with each ring of picking 1 of oese, is respectively connected to equipped with 100mL (1 bottle is respectively connect in the triangular flask of potato nutrient solution), 2d is cultivated in 30 DEG C.Taken from candida utili nutrient solution again 8mL and from 2mL combined inoculations are taken in kluyveromyces marxianus nutrient solution in 1000mL potato nutrient solutions, 2d is cultivated in 30 DEG C, must mixed Yeast juice.Take again compound barm liquid 10g access 1000g wheat bran corn culture mediums in (︰ 8 of bran skin ︰ jade rice ︰ water=8 ︰ 2, in advance prior to Sterilize 30min at 121 DEG C, standby after cooling), after being well mixed, make thinner in the ceramic whiteware disk for sterilizing, thickness about 10mm, 2d is cultivated in 30 DEG C, is compound barm solid spawn；

C, prepare nutritive salt：Take 4000g KH₂PO4、3200g KCl、800gMgSO₄、200gFeSO₄And 100gZnSO₄, Crush mixing standby；

D, prepare mixing enzyme preparation：By phytase 1000g, cellulase 100g, zytase 50g, pectase 100g and Papain 50g mixes standby；

E, Rapeseed Meal by Aspergillus Fermentation：Rapeseed dregs and corn are crushed with the pulverizer of 2.0mm sieve apertures, rapeseed dregs 980kg, corn flour is taken 5kg, nutritive salt 2.5kg, mixing enzyme preparation 0.5kg, mixed yeast solid spawn 12kg, bacillus licheniformis bacterium solution 2kg and Water 898kg, after all raw materials are mixed, loads self-control stainless steel case, is paved into thickness 500mm, after capping, 20h is cultivated in 30 DEG C, Then compost in case is poured into self-control stainless steel tray, is paved into thickness 40mm, continue to cultivate 35h in 30 DEG C, take out nature It is rapeseed peptide (rsp) albumen finished product after air-drying.

Each composition of gained finished product is：Crude protein 44%, total amino acid content improves 16%, giucosinolate clearance 80%, phytic acid degradation rate 45%, pectin degrading rate 40%, acid-soluble protein content increases by 100%.

Feed experiment：40kg Sichuan Local Black Pigs (Neijiang Pig) is carried out with the present embodiment rapeseed peptide (rsp) albumen formula feed 30d growth performances are determined, and rapeseed peptide (rsp) albumen is accounted for 12% in complete diet pellet, without antibiotic, its complementary energy and protein level with compare Group is consistent (control group is added with antibiotic).Result shows：Experimental group and control group do not occur dead and have loose bowels situation, experimental group Daily gain reaches 860g, more than control group 8.8%, so that yuan/pig of reduces cost 18.

Nursing experiment of the preparation and product of the rapeseed peptide (rsp) albumen of embodiment 2 to pig

Each method for preparing raw material of the present embodiment is carried out according to the method for embodiment 1.

Crush rapeseed dregs with the pulverizer of 2.0mm sieve apertures, take rapeseed dregs 990kg, nutritive salt 4kg, mixing enzyme preparation 1kg, Water 900kg, bacillus licheniformis bacterium solution 1kg, mixed yeast solid spawn 5kg, after all raw materials are mixed, load self-control not Rust steel case, is paved into thickness 500mm, after capping, 20h is cultivated in 30 DEG C, and compost in case then is poured into self-control stainless steel tray In, thickness 30mm is paved into, continue to cultivate 35h in 35 DEG C, it is rapeseed peptide (rsp) albumen finished product after taking-up natural air drying.

Each composition of gained finished product is：Crude protein 44%, giucosinolate clearance 82%, phytic acid degradation rate 48%, pectin Degradation rate 30%, acid-soluble protein content increases by 90%.

Feed experiment：With the present embodiment rapeseed peptide (rsp) albumen formula feed, 60kg growing and fattening pigs (Sichuan Local Black Pigs) are carried out 30d performance measurements, wherein, control group is accounted for 20% in complete diet pellet for soybean meal content；Experimental group is accounted in complete diet pellet for rapeseed peptide (rsp) albumen 16%, dregs of beans 5%, without antibiotic, its complementary energy is consistent with control group with protein level experimenter.

Result shows：There was no significant difference with control group weightening for experimental group, but experimental group hair color light is better than control group, Sulfur amino acid content is higher than dregs of beans in being attributable to rapeseed peptide (rsp) albumen；Interior during feeding 30d, experimental group is raised than control group reduction Material yuan/the pig of cost 11.

Nursing experiment of the preparation and product of the rapeseed peptide (rsp) albumen of embodiment 3 to pig

Rapeseed dregs and corn are crushed with the pulverizer of 2.0mm sieve apertures, rapeseed dregs 980kg, corn flour 8kg, nutritive salt is taken 2.5kg, mixing enzyme preparation 0.5kg, water 900kg, bacillus licheniformis bacterium solution 1kg, mixed yeast solid spawn 9kg, by institute After thering is raw material to mix, load self-control stainless steel case, be paved into thickness 500mm, after capping, 24h is cultivated in 28 DEG C, then will be trained in case Nutriment is poured into self-control stainless steel tray, is paved into thickness 40mm, continues to cultivate 40h in 28 DEG C, is vegetable seed after taking-up natural air drying Peptide albumen finished product.

Each composition of gained finished product is：Crude protein 42%, giucosinolate clearance 75%, phytic acid degradation rate 35%, pectin Degradation rate 32%, acid-soluble protein content increases by 80%.

Feed experiment：With the present embodiment rapeseed peptide (rsp) albumen formula feed, antibiotic-free is carried out to this lattice pig system and feeds experiment. 20 piglets of control group feed alfalfa extracts nonreactive feed from 30kg；20 piglets of experimental group feed rapeseed peptide (rsp) albumen nonreactive from 30kg (30kg~75kg stages, rapeseed peptide (rsp) albumen accounts for complete diet pellet 9% to feed；75kg~120kg fattening stages, rapeseed peptide (rsp) albumen is accounted for entirely Valency material 13%), experimental group and control group are not added with any antibiotic, deliver weight 120 kg for sale.

Result shows：There was no significant difference for experimental group and control group growth, does not occur sufferer, and whole feedstuff-meat ratio is 2.7, so that experimental group feed cost 25 yuan/pigs of reduction.With reference to green food standard detection, 30 indexs of censorship all reach To green food standard.The mechanism of rapeseed peptide (rsp) albumen nonreactive cultivation is that, containing a large amount of probiotics, can improve animal body in feed Nospecific immunity and intestinal health level.

Nursing experiment of the preparation and product of the rapeseed peptide (rsp) albumen of embodiment 4 to pig

Rapeseed dregs and corn are crushed with the pulverizer of 2.0mm sieve apertures, rapeseed dregs 980kg, corn flour 5kg, nutritive salt is taken 3kg, mixing enzyme preparation 1kg, water 900kg, bacillus licheniformis bacterium solution 1.5kg, mixed yeast solid spawn 11kg, will be all After raw material is mixed, load self-control stainless steel case, be paved into thickness 450mm, after capping, 20h is cultivated in 32 DEG C, then will be cultivated in case Material is poured into self-control stainless steel tray, is paved into thickness 40mm, continues to cultivate 35h in 30 DEG C, is rapeseed peptide (rsp) after taking-up natural air drying Albumen finished product.

Each composition of gained finished product is：Up to 43.5%, total amino acid content improves 16%, giucosinolate removal to crude protein Rate 80%, phytic acid degradation rate 45%, pectin degrading rate 40%, acid-soluble protein content increases by 100%.

Feed experiment：With the present embodiment rapeseed peptide (rsp) albumen formula feed, nonreactive nursing is carried out to native di (hetero) pig (white hair pig) And rapeseed peptide (rsp) albumen threshold dose experiment (observation has non-toxic reaction and growth performance).Experiment is divided to two groups, every group 10, reality Test group one：Rapeseed peptide (rsp) albumen consumption accounts for complete diet pellet 19%；Experimental group two：Rapeseed peptide (rsp) albumen consumption accounts for complete diet pellet 27%；Control group： Plant's corn-soybean meal is from dispensing, and each group energy nitrogen nutrition level is consistent, two experimental groups not added with antibiotic, from 35kg Feed 27d.

Result shows：Total augment weight：Control group 173.4kg, the 186.6kg of experimental group one, the 158.6kg of experimental group two；Feedstuff-meat ratio： Control group 2.996, experimental group 1, experimental group 2 2.985；Experimental group one grows fastest, compared to control group, can save About 0.056 yuan/kg of cost (weightening), quickly being delivered for sale if desired for pig can select this formulation material；The speed of growth of experimental group two is most slow, But cost-effective obvious, cost-effective 0.7122 yuan/kg (weightening), if growth cycle need to be extended, can use this formulation material.Respectively Group does not occur sufferer, and feed intake is not apparent from declining (good palatability).Result above shows that rapeseed peptide (rsp) albumen consumption accounts for complete diet pellet 27%, pig is in a good state of health, and (common unprocessed rapeseed dregs consumption is 5%, it may appear that poison to have no the toxic reaction of rapeseed dregs Property reaction).

Nursing experiment of the preparation and product of the rapeseed peptide (rsp) albumen of embodiment 5 to chicken

Rapeseed dregs and corn are crushed with the pulverizer of 2.0mm sieve apertures, rapeseed dregs 980kg, corn flour 5kg, nutritive salt is taken 2.2kg, mixing enzyme preparation 0.8kg, water 900kg, bacillus licheniformis bacterium solution 2kg, mixed yeast solid spawn 12kg, by institute After thering is raw material to mix, load self-control stainless steel case, be paved into thickness 500mm, after capping, 18h is cultivated in 30 DEG C, then will be trained in case Nutriment is poured into self-control stainless steel tray, is paved into thickness 40mm, continues to cultivate 30h in 33 DEG C, is vegetable seed after taking-up natural air drying Peptide albumen finished product.

Each composition of gained finished product is：Up to 43%, total amino acid content improves 16%, giucosinolate clearance to crude protein 75%, phytic acid degradation rate 40%, pectin degrading rate 35%, acid-soluble protein content increases by 100%.

Feed experiment：Chicken feed is prepared with the present embodiment rapeseed peptide (rsp) albumen, growth performance and premunition is observed.From 300 age in days of health 1 " Daheng 199 " cocks, are randomly divided into 2 treatment, each 3 repetition for the treatment of, and each repeats 50 chickens.It is right It is corn-soybean meal basal diet according to group, adds 21% rapeseed peptide (rsp) albumen respectively in daily ration based on treatment group and carry out Substitution for Soybean Meal, After raising 30d, random 20 broiler chicken of extraction are weighed on an empty stomach from each column, calculate broiler chicks daily gain, feedstuff-meat ratio, feed intake, profit Analyzed with SPSS13.0 software statistics.

Result shows：Compared with control group corn-soybean meal diet, experimental group daily ingestion amount, daily gain, feedstuff-meat ratio etc. are raw Performance long does not make significant difference, see the table below, the low 0.45 yuan/kg of experimental group per gain weight feed cost, reduces 7.2%, and Due to antibiotic-free addition in daily ration, the survival rate of broiler chicks improves 3%.

	Control group	Experimental group
			Experiment initial weight (kg)	0.04	0.04
Experiment end weight (kg)	0.72±0.11	0.71±0.09
			Daily ingestion amount (g/d)	45.6±0.03a	46.2±0.05a
Daily gain (g/d)	22.7±0.01	22.3±0.01
			Feed conversion rate	2.01±0.03a	2.07±0.06a

Note：Back of the body mark is identical to represent difference not significantly (p ＞ 0.05)

Nursing experiment of the preparation and product of the rapeseed peptide (rsp) albumen of embodiment 6 to chicken

Rapeseed dregs and corn are crushed with the pulverizer of 2.0mm sieve apertures, rapeseed dregs 980kg, corn flour 10kg, nutritive salt is taken 2.5kg, mixing enzyme preparation 0.5kg, water 900kg, bacillus licheniformis bacterium solution 1.5L, mixed yeast solid spawn 7kg, by institute After thering is raw material to mix, load self-control stainless steel case, be paved into thickness 450mm, after capping, 20h is cultivated in 30 DEG C, then will be trained in case Nutriment is poured into self-control stainless steel tray, is paved into thickness 50mm, continues to cultivate 35h in 30 DEG C, is vegetable seed after taking-up natural air drying Peptide albumen finished product.

Each composition of gained finished product is：Crude protein 42.5%, total amino acid content improves 16%, giucosinolate clearance 75%, phytic acid degradation rate 35%, pectin degrading rate 35%, acid-soluble protein content increases by 100%.

Feed experiment：Small middle broiler fodder is prepared with the present embodiment rapeseed peptide (rsp) albumen, growth performance and premunition is observed.Choosing With Daheng's broiler chicken 300, it is divided into two groups, test group 180, control group 120.Feed in two stages：1st stage went out shell baby chick Cultivate to 33 ages in days, the 2nd stage is from 33 ages in days to 44 ages in days.The stage of control group the 1st uses honest commercial feed, and the 2nd stage is with newly Wish commercial feed；The stage feed of experimental group the 1st, rapeseed peptide (rsp) albumen dosage is 12%, and the 2nd stage feed, rapeseed peptide (rsp) albumen adds Measure is 16%.Result such as following table：

1st stage：Experimental group often increases by 1 jin of meat, 0.5 yuan of feed cost reduction, the 2nd stage：Experimental group often increases by 1 jin Meat, 1 yuan of feed cost reduction does not add antibacterials in experimental group feed, whole dead substantially suitable with control group.

Nursing experiment of the preparation and product of the rapeseed peptide (rsp) albumen of embodiment 7 to chicken

Rapeseed dregs and corn are crushed with the pulverizer of 2.0mm sieve apertures, rapeseed dregs 980kg, corn flour 7kg, nutritive salt is taken 2.5kg, mixing enzyme preparation 0.5kg, water 900kg, bacillus licheniformis bacterium solution 2kg, mixed yeast solid spawn 10kg, by institute After thering is raw material to mix, load self-control stainless steel case, be paved into thickness 500mm, after capping, 20h is cultivated in 30 DEG C, then will be trained in case Nutriment is poured into self-control stainless steel tray, is paved into thickness 60mm, continues to cultivate 32h in 32 DEG C, is vegetable seed after taking-up natural air drying Peptide albumen finished product.

Each composition of gained finished product is：Crude protein 42.8%, total amino acid content improves 16%, giucosinolate clearance 75%, phytic acid degradation rate 35%, pectin degrading rate 40%, acid-soluble protein content increases by 95%.

Feed experiment：Big broiler fodder, observes growth performance and premunition in being prepared with the present embodiment rapeseed peptide (rsp) albumen.Choosing With the choice meat of healthy weight 0.8kg or so with the white skin fiber crops plumage cock of " Daheng 199 " green grass or young crops pin, 3 treatment are randomly divided into, at each 3 repetitions of reason, each repeats 50.Control group is with corn soybean diet (added with antibiotic)；Experimental group is divided into two nutritive waters It is flat, equal not added with antibiotic：In the feed of experimental group one, rapeseed peptide (rsp) albumen addition 25%, in the feed of experimental group two, rapeseed peptide (rsp) albumen adds Plus 24%, feed phase 60d.The growth performances such as chicken daily gain, feedstuff-meat ratio, feed intake are calculated in units of circle, and determines pigeon breast Meat amino acid index and normal blood index, use SPSS13.0 software analysis, as a result see the table below：

	Control group	Experimental group 1	Experimental group 2
				Experiment initial weight (kg)	0.78±0.12	0.77±0.09	0.76±0.11
Experiment end weight (kg)	2.53±0.09	2.51±0.07	2.49±0.05
				Daily ingestion amount (g/d)	752.5±5.46	757.0±2.65	764.0±4.31
Daily gain (g/d)	292±1.87	290±2.25	283±4.07
				Feed conversion rate (FCR)	2.58±0.04a	2.61±0.02ab	2.65±0.03b

Pigeon breast muscle amino acid index analysis shows, experimental group one, compared with control group, amino acid content is total in its muscle Amount without significant difference, respectively 56.63% and 57.77%, wherein the equal nothings of Glu, Gly, Asp, Ala and Arg related to delicate flavour Significant difference；And experimental group two, in pigeon breast muscle 5 kinds of Fresh ear field contents and total amino acid content be respectively 27.33% and 60.86%, control group 5.1% and 7.5% is higher by respectively.

The routine blood indexes initial analysis of chicken：(1) lymphocyte rate：Control group, experimental group one and experimental group two are respectively： 59.7%th, 72.05% and 73.05%；(2) total white blood cells (WBC)：Control group, experimental group one and experimental group two are respectively： 100.8th, 105.3 and 121.4 (unit is：×10¹²/L)。

Result above shows：(1) rapeseed peptide (rsp) albumen alternative most of dregs of beans in big daily grain of chicken in, produces a high-quality Broiler chicken can increase 2.5 yuan~3.2 yuan of profit newly；(2) experimental group is in two blood of reflection immunity of organisms of lymphocyte rate and WBC It is better than control group on conventional index；(3) prompting is by Tiny ecosystem nutrition technique controllable livestock products local flavor index and improves livestock products Product quality.

Nursing experiment of the preparation and product of the rapeseed peptide (rsp) albumen of embodiment 8 to milk cow

Rapeseed dregs and corn are crushed with the pulverizer of 2.0mm sieve apertures, rapeseed dregs 990kg, corn flour 3kg, nutritive salt is taken 2.5kg, mixing enzyme preparation 0.5kg, water 900kg, bacillus licheniformis bacterium solution 1.5kg, mixed yeast solid spawn 4kg, will After all raw materials are mixed, load self-control stainless steel case, be paved into thickness 400mm, after capping, 24h is cultivated in 30 DEG C, then by case Compost is poured into self-control stainless steel tray, is paved into thickness 20mm, continues to cultivate 35h in 32 DEG C, is dish after taking-up natural air drying Seed peptide albumen finished product.

Each composition of gained finished product is：Crude protein 43.1%, total amino acid content improves 16%, giucosinolate clearance 75%, phytic acid degradation rate 35%, pectin degrading rate 35%, acid-soluble protein content increases by 90%.

Feed experiment：Milk cow forage is prepared with the present embodiment rapeseed peptide (rsp) albumen.From lactation period Fresian 10, point It is control group and experimental group, each 5.Control group former cattle farm from batch formula；Nutrition of the experimental group with reference to control group of formula Level, rapeseed peptide (rsp) albumen accounts for 15% in group of formula is tested.Two groups of feed formula cost price is identical.Fed through 30d, two groups of products The butterfat percnetage of milk is identical, and the experimental group output of milk is higher than control group 6.5%.

Nursing experiment of the preparation and product of the rapeseed peptide (rsp) albumen of embodiment 9 to sheep

Rapeseed dregs and corn are crushed with the pulverizer of 2.0mm sieve apertures, rapeseed dregs 985kg, corn flour 4kg, nutritive salt is taken 2.5kg, mixing enzyme preparation 0.5kg, water 900kg, bacillus licheniformis bacterium solution 1.5kg, mixed yeast solid spawn 8kg, will After all raw materials are mixed, load self-control stainless steel case, be paved into thickness 400mm, after capping, 20h is cultivated in 30 DEG C, then by case Compost is poured into self-control stainless steel tray, is paved into thickness 45mm, continues to cultivate 36h in 30 DEG C, is dish after taking-up natural air drying Seed peptide albumen finished product.

Each composition of gained finished product is：Crude protein 43%, total amino acid content improves 16%, giucosinolate clearance 80%, phytic acid degradation rate 37%, pectin degrading rate 40%, acid-soluble protein content increases by 90%.

Feed experiment：Sheep feed is prepared with the present embodiment rapeseed peptide (rsp) albumen.The big ear sheep ewe in Jianyang 20 is selected, two are divided at random Group.Control group feed：Rye grass 70%, corn 10%, dregs of beans 10%, wheat bran 10%；Experimental group feed：Rye grass 70%, jade Rice 10%, rapeseed peptide (rsp) protein 12 %, wheat bran 8%, about 210 yuan of experimental group concentrated feed reduces cost per ton are fed through 30d, experiment Group average growth rate is higher than control group 5% or so.

Comparative example 1

Rapeseed dregs 950kg, the wheat bran 40kg and adjusting material 10kg after crushing are weighed, it is another to prepare triangular flask bacterial liquid 3L.Its In, the adjusting material is by 0.1%KH₂PO₄, 0.8% urea, 0.1%MgSO₄, 0.2%CuSO₄.5H₂O and corn flour mixing group Into rapeseed dregs moisture is 10%.Per 100kg, above-mentioned material adds strain (candida tropicalis Candidatropicalis) liquid 300mL and clean water 40kg are stirred, sabot, and heat and moisture preserving is completed in 30 DEG C of fermentations, 48h after fermentation.

	Comparative example	Rapeseed peptide (rsp) albumen of the present invention
			Rapeseed dregs accounts for fermentation material ratio	94~96	97~99
Major auxiliary burden and consumption	Wheat bran 3~4%	Corn flour≤1.5%
			Fermented bacterium	1 kind (candida tropicalis)	3 kinds
Complex enzyme is used	Nothing	5 kinds
			Zymotechnique	Aerobic fermentation	Anaerobic fermentation (enzymolysis)+aerobic fermentation
Sulphur glucoside clearance	65%	75~82%
			Remove the ancillary method of sulphur removal glucoside	Plus CuSO₄·5H₂O	Need not
Crude protein	Up to 44%	42.5~44%
			Total amino acid is improved	Be not given	16%
Phytic acid degradation rate	Be not given	35~45%
			Pectin degrading rate	Be not given	30~40%
The molten albumen increment rate of acid	Be not given	90~100%

In summary it can be seen, by the effect of the raw materials such as bacterial strain in inventive formulation and enzyme preparation, make the sulphur in rapeseed dregs Clearance, phytic acid degradation rate, pectin degrading rate of glycoside etc. are obtained for very big raising；So that the rapeseed peptide (rsp) for preparing Albumen can be applied in the raising of livestock and poultry well, and achieves high-quality, the effect of low cost.So, this The formula and method of invention treatment rapeseed dregs provide a more preferable approach for effective utilization of rapeseed dregs.

Claims

1. rapeseed peptide (rsp) albumen, it is characterised in that：Formed by the fermenting raw materials of following weight proportion：97~99 parts of rapeseed dregs powder, jade 0~1.5 part of ground rice, 0.4~1.5 part of mixed yeast solid spawn, 0.1~0.4 part of nutritive salt, mixing enzyme preparation 0.03~ 0.1 part, 0.1~0.3 part of Bacillus licheniformis liquid, water；Wherein, the gross weight of bacillus licheniformis bacterium solution and water account for other five Plant raw material gross weight 0.6~1.2 times；Wherein, described rapeseed dregs is 95 type rapeseed dregs；

The preparation method of described mixed yeast solid spawn is：From the oblique of candida utili bacterium (Candida utilis) During picking strain accesses potato nutrient solution in the preservation of bacteria strain of face, in 28~32 DEG C of temperature 2~3d of culture, candida utili is obtained Bacteria culture fluid；The picking bacterium from the slant preservation strain of kluyveromyces marxianus bacterium (Kluyveromyces marxianus) Plant in access potato nutrient solution, in 28~32 DEG C of temperature 2~3d of culture, obtain kluyveromyces marxianus bacteria culture fluid；Press again Weight ratio, candida utili bacteria culture fluid:Kluyveromyces marxianus bacteria culture fluid=0.75~0.9:0.1~0.25 mixing Access in potato nutrient solution, in 28~32 DEG C of temperature 2~3d of culture, that is, obtain compound barm liquid；Compound barm liquid is transferred to again In 28~32 DEG C of temperature 2~3d of culture in wheat bran corn culture medium, mixed yeast solid spawn is obtained final product；Marx's Crewe The deposit number for tieing up saccharomycete is CGMCC No.9427；

The nutritive salt includes following component：1 part of KH is counted by weight ratio₂PO₄, 0.8 part of KCl, 0.2 part of MgSO₄, 0.05 part FeSO₄With 0.025 part of ZnSO₄；

The mixing enzyme preparation includes following component：Count by weight ratio, phytase:Cellulase:Zytase:Pectase: Papain=8~12:0.5~1.5:0.3~0.8:0.8~1.5:0.4~0.6；

The preparation method of described bacillus licheniformis bacterium solution is：From bacillus licheniformis (Bacillus licheniformis) Slant preservation strain in picking strain access meat soup protein culture medium in, 34~38 DEG C cultivate 30~40h, obtain final product lichens Bacillus bacterium solution；

The preparation method of the rapeseed peptide (rsp) albumen, comprises the following steps：

B, fermentation：After fermentation material is fermented into 18~24h in 28~32 DEG C under the anaerobic state, take out, then under aerobic state in Fermented 30~40h at 28~35 DEG C, and rapeseed peptide (rsp) albumen is obtained after drying.

2. rapeseed peptide (rsp) albumen according to claim 1, it is characterised in that：Formed by the fermenting raw materials of following weight proportion： 98 parts of rapeseed dregs powder, 0.5 part of corn flour, 1.2 parts of mixed yeast solid spawn, 0.25 part of nutritive salt, mixing enzyme preparation 0.05 Part, 0.15 part of Bacillus licheniformis liquid, water；Wherein, bacillus licheniformis bacterium solution accounts for other five kinds of raw materials with the gross weight of water 0.9 times of gross weight.

3. rapeseed peptide (rsp) albumen according to claim 1, it is characterised in that：The candida utili bacteria culture fluid:Mark This kluyveromyces bacteria culture fluid=0.85:0.15.

4. the preparation method of the rapeseed peptide (rsp) albumen described in any one of claims 1 to 3, it is characterised in that：Comprise the following steps：

5. the preparation method of rapeseed peptide (rsp) albumen according to claim 4, it is characterised in that：In step B, anaerobic state fermentation When fermentation material thickness be 400~500mm；The thickness of fermentation material is no more than 60mm during aerobic state fermentations.