CN109757610A - Fish special feed cooperates additive - Google Patents
Fish special feed cooperates additive Download PDFInfo
- Publication number
- CN109757610A CN109757610A CN201811374998.6A CN201811374998A CN109757610A CN 109757610 A CN109757610 A CN 109757610A CN 201811374998 A CN201811374998 A CN 201811374998A CN 109757610 A CN109757610 A CN 109757610A
- Authority
- CN
- China
- Prior art keywords
- fish
- protease
- special feed
- iii
- oligosaccharides
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Fodder In General (AREA)
- Feed For Specific Animals (AREA)
Abstract
The present invention provides fish special feed and cooperates additive, belongs to fish culture technical field, including (I) oligosaccharide composition；And/or (II) carboxylate；(III) proteinase complex；(III) proteinase complex is by cultivating the proteinase complex comprising effective proteinase mixture dredging continuous shape hyphomycete and obtaining.The present invention also provides a kind of fish special feeds for cooperating additive containing basal feed and fish special feed.Have the beneficial effect that fish special feed cooperation additive helps to improve fish intestinal flora and can promote intestinal villus regeneration, improve intestinal absorption efficiency, it maintains gastrointestinal health and/or reduces the demand to antibiotic, reduce the abuse of antibiotic in breeding process, hence it is evident that reduce the morbidity and mortality of fish.
Description
Technical field
The invention belongs to fish culture technical fields, and in particular to fish special feed cooperates additive.
Background technique
Fish is Perciformes Sciaenidae, and body colour is silver grey.Voluminous East Sea Zhoushan ocean surface.Fish generally every urosome is long by 45~
55cm, 500~1000g of weight, maximum have over one hundred jin.It is distributed in the waters such as Bohai and Yellow Seas, the East Sea, the South Sea.Fish body extend and
Flat-sided, back, the shallow arc of abdomen, big individual is big up to more than 10 kilogram, mouthfuls and slightly-inclined, body are tiny by ctenoid scale, scale, and surface layer is thick
Rough, head is arranged by 4, cycloid scale, jaw hole, central jaw hole and inside jaw hole in square, no jaw palpus.Upper jaw layman tooth and lower jaw
Expert's tooth expands, in toothlike, inner face denticle become band group, eyelet is big, eye mask transparency is high red and bright.Body is in bluish grey brown
Color, abdomen be canescence, 2, dorsal fin connect together, centre has one deep to incise.Fin spine upper limb is black, and fin ray portion center has one
Stringer blackstreak；Pectoral fin base portion yellow, edge black；Anal fin has 2 spine；Tail fin is wedge shaped, and the base portion of tail fin is yellow, side
Edge color is slightly shallow.
Piscinity is sweet, salty, flat, has blood-nourishing, stops blooding, invigorates the kidney and stops nocturnal emission and anti-inflammatory effect, to treatment alpastic anemia, spits
Blood, emission due to the kidney deficiency, pain swollen, nameless sores or boils, mazoitis etc. are effectively.Flesh of fish delicious is one of important marine products economic fish,
It is the higher fish of economic value, thus is liked deeply by consumer and culturist.At present due to artificial high-density breeding fish,
The immunity of fish, anti-stress ability relative drop, are easy to appear lesion, cause large stretch of death, seriously affect aquaculture economy.
In aquaculture industry, in order to improve cultured output and benefit, often using antibiotic prophylaxis and treatment fish disease, some is also
Promote fish growth using antibiotic and improves the utilization rate of fish feed.But the abuse of antibiotic while being good for for aquatic livestock
The edible safety of Kang Chengchang and the mankind have buried deep hidden danger, and abuse of antibiotics can not be ignored at present.
Summary of the invention
The purpose of the present invention is to provide fish special feeds to cooperate additive, helps to improve fish intestinal flora and can
Promote intestinal villus regeneration, improve intestinal absorption efficiency, maintain gastrointestinal health and/or reduce the demand to antibiotic, reduces cultivation
The abuse of antibiotic in the process, hence it is evident that reduce the morbidity and mortality of fish.
The technical solution that the present invention is taken to achieve the above object are as follows:
The present invention provides a kind of fish special feed cooperation additive comprising:
(I) oligosaccharide composition；And/or
(II) carboxylate；With
(III) proteinase complex；(III) proteinase complex is by cultivating the packet dredging continuous shape hyphomycete and obtaining
Proteinase complex containing effective proteinase mixture.Fish special feed cooperation additive helps to improve the micro- life of fish enteron aisle
Object flora, maintains gastrointestinal health and/or reduction pair at the intestinal absorption for promoting the regeneration of fish intestinal villus, improving purpose nutriment
The demand of antibiotic reduces the abuse of antibiotic in breeding process, can also significantly reduce feedstuff-meat ratio when fish culture, shortens and supports
The period is grown, in addition, above-mentioned proteinase complex helps to strengthen the immune system of fish, can obviously reduce and caused by germ contamination
Fish morbidity and mortality, be a kind of efficient fish special feed cooperation additive.
(I) oligosaccharide composition is distributed with following glycosidic bond: has and is no less than 8mol% α -1,6 glycosidic bonds, and not
Less than 8mol% β -2,1 glycosidic bond, and/or no less than 8mol% β -1,3 glycosidic bonds, and wherein at least 8 dry weight % oligosaccharides groups
The degree of polymerization for closing object is 2~6.
(I) oligosaccharide composition includes: isomaltose-oligosaccharides, isomaltose-inulobiose-oligosaccharides, two pool of inulin-
Oligosaccharides, two pools of chondroitin-oligosaccharides, two pools of hyalomitome-oligosaccharides, two pools of hyalomitome-isomaltose-oligosaccharides, two pool of chondroitin-different wheat
Bud sugar-oligosaccharides, galacto-oligosaccharide, lactose-two pools of hyalomitome-oligosaccharides, isomaltose-galacto-oligosaccharide, lactose-inulobiose-widow
Sugar, inulobiose-isomaltose-oligosaccharides.Oligosaccharide composition is applied in fish feed additive special or feed to feed
Fish has beneficial health benefit, helps to reduce the fish death rate, improves intestinal microflora, improves nutriment suction
The demand to antibiotic is received, maintains gastrointestinal health and/or reduced, the abuse of antibiotic in breeding process is reduced.
(II) carboxylate is selected from:
According to the lactoyl carboxylate of formula 1,
Formula 1:R1-COO-[-CH(CH3)-COO]n-R2
Or/and its Ba, Be, Mn (II), Ca, Na, NH4Or Co (II) salt,
According to the glycollyl carboxylate of formula 2,
Formula 2:R1-COO-[-CH2-COO]n-R2
Or/and its Ba, Be, Mn (II), Ca, Na, NH4Or Co (II) salt,
In above formula, R1Selected from chlorphenyl or bromophenyl, R2Selected from C2~C10Alkyl or alkenylene chain, and n indicates number
The integer that value is 1~10；Above-mentioned carboxylate products application can significantly increase in fish special feed or feed addictive
The antibacterial activity of fish, has the function of beneficial in the prevention and treatment of fish enteric infection, and helps to improve
The growth rate of fish reduces feedstuff-meat ratio and culture-cycle, increases economic benefit.
(III) proteinase complex is to be mixed by what the thin continuous shape hyphomycete of culture obtained comprising effective protease
The proteinase complex of object, the ratio of effective proteinase mixture are more than active the 75% of the proteinase complex,
In the protease of effective proteinase mixture, a kind of protease has 7.4 or so isoelectric point, and another protease tool
There is 8.3 or so isoelectric point；Proteinase complex peomotes fish such as fish intestinal villus and regenerates, and therefore greatly changes
The intestinal absorption of kind purpose nutriment, so that feedstuff-meat ratio when significantly reducing fish culture, shortens the culture-cycle, in addition, above-mentioned
Proteinase complex helps to strengthen the immune system of fish, can obviously reduce the disease incidence of the fish as caused by germ contamination,
The especially death rate.
Preferably, in (III) proteinase complex isoelectric point be 7.4 or so protease have 82000~
90000 Anson units/mg protein specific activity.
Preferably, in (III) proteinase complex isoelectric point be 8.3 or so protease have 20000~
25000 Anson units/mg protein specific activity.
Preferably, (III) proteinase complex is used for fish special feed as the supplement of feed addictive
In, the feed addictive exists with powder, liquid or other any suitable forms mixed with fodder compound.
Preferably, cultivating to obtain (III) proteinase complex and dredging continuous shape hyphomycete bacterial strain and filtering fermentating liquid, so
After set about carrying out extracting proteinase complex by ultrafiltration and isoelectric focusing, finally proteinase complex obtained is lyophilized
It crushes, specifically includes:
Fermentation: single colonie (the decomposable culture of the produced protease of bacterial strain of transparent circle is selected from primary dcreening operation culture medium flat plate
Substrate protein in base, therefore transparent circle phenomenon is presented in periphery of bacterial colonies), scribing line separation；Bacterial strain after purification, which is forwarded to, fills fermentation
In the fermentation shake flask of culture medium, produced in filtrated air/culture volume/minute ventilation of 0.3 volume, and in fermentation
4~6d of ferment at constant temperature under the conditions of enzyme medium pH is 7.2~7.5, temperature is 40~45 DEG C, revolving speed is 150~200r/min,
Supernatant is taken after centrifugation；
Ultrafiltration: ultrafiltration is carried out by means of the film with 50kDa cutoff threshold, and ultrafiltration carries out 3~10 hours；
Isoelectric focusing: controlling solution ph respectively is 7.4 ± 0.1 and 8.3 ± 0.1 progress isoelectric focusing, i.e. acquisition accounts for more
Several protease with 7.4 or so isoelectric points (pI), and with less ratio, the albumen with 8.3 or so isoelectric points (pI)
Enzyme；Up to (III) proteinase complex after freeze-drying crushing.
As to present invention further optimization, enzymatic production culture medium includes the substance of following weight content: 180~
240 mesh Corncob Xylans 0.18~0.2%, threitol 0.012~0.014%, tryptone 1.8~3%, yeast extract
0.3~0.5%, sodium nitrate 0.1~0.2%, KH2PO40.5~0.6%, K2HPO40.15~0.2%, MgSO40.03~
0.05%, natural pH；The carbon source of particular particle size facilitates the output of high enzyme activity protease, appropriate particle size in enzymatic production culture medium
Carbon source can make microorganism own growth procreation with shaking flask in dissolved oxygen amount formed balance, also can avoid carbon source particle it is meticulous and with
Other nutriments form the biggish suspension of viscosity and influence the generation of Metabolite in turn, and it is horizontal to reduce producing enzyme；This
Outside, the Corncob Xylan and threitol that special proportion is added in enzymatic production culture medium have the output of high enzyme activity protease
Stimulation is induced, can greatly accelerate the secretion for dredging the highly active protein enzyme that continuous shape hyphomycete bacterial strain is 7.4 or so to isoelectric point
With precipitation, to significantly improve the albumen that isoelectric point is 7.4 or so on the basis of maintaining the content of whole proteinase complex
The accounting of enzyme improves the activity of proteinase complex on the whole.
It is dedicated containing basal feed and fish in the fish special feed the present invention also provides a kind of fish special feed
Feed formulation additive,
The basal feed includes:
The protein of 31~35wt%；
The fat of 22~24wt%；
The nucleotide and/or nucleosides of 0.06~0.09wt%；
The digestible sulfur-containing amino acid of 2.3~5.0wt%；
Total effective calcium of 0.3~0.5wt%；
Total available phosphorus of 0.32~0.4wt%；
The sodium of 0.3~0.35wt%.
The fish special feed cooperation additive includes:
(I) oligosaccharide composition；And/or
(II) carboxylate；With
(III) proteinase complex；(III) proteinase complex is by cultivating the packet dredging continuous shape hyphomycete and obtaining
Proteinase complex containing effective proteinase mixture.
The invention has the benefit that
1) oligosaccharide composition is applied in fish feed additive special or feed feeds fish with beneficial health
Effect helps to reduce the fish death rate, improves intestinal microflora, improve absorption of nutrient ingredients, maintain gastrointestinal health
And/or demand of the reduction to antibiotic, reduce the abuse of antibiotic in breeding process；
2) carboxylate products application can significantly increase the antibacterium of fish in fish special feed or feed addictive
Activity has the function of growth speed that is beneficial, and helping to improve fish in the prevention and treatment of fish enteric infection
Rate reduces feedstuff-meat ratio and culture-cycle, increases economic benefit；
3) proteinase complex peomotes fish such as fish intestinal villus and regenerates, and therefore dramatically improves purpose battalion
The intestinal absorption of substance is supported, so that feedstuff-meat ratio when significantly reducing fish culture, shortens the culture-cycle, in addition, above-mentioned protease is multiple
Closing object helps to strengthen the immune system of fish, can obviously reduce the disease incidence of the fish as caused by germ contamination, especially extremely
Die rate；
4) when fermentation obtains proteinase complex, the carbon source that particular particle size is added in enzymatic production culture medium facilitates height
The output of enzyme activity protease, it is flat that the carbon source of appropriate particle size can be such that the procreation of microorganism own growth is formed with the dissolved oxygen amount in shaking flask
It weighs, also avoidable carbon source particle is meticulous and simultaneously influence bacterial strain is metabolized in turn with other nutriments formation biggish suspension of viscosity
It is horizontal to reduce producing enzyme for the generation of product；
5) Corncob Xylan and threitol that special proportion is added in enzymatic production culture medium are to high enzyme activity protease
Output has induction stimulation, can greatly accelerate and dredge the highly active protein enzyme that continuous shape hyphomycete bacterial strain is 7.4 to isoelectric point
Secretion and precipitation, to significantly improve accounting for for highly active protein enzyme on the basis of maintaining the content of whole proteinase complex
Than improving the activity of proteinase complex on the whole.
Fish special feed is provided present invention employs above-mentioned technical proposal and cooperates additive, compensates for the prior art not
Foot, reasonable design, easy operation.
Specific embodiment
Present invention is further described in detail with reference to embodiments:
Embodiment 1:
A kind of fish special feed cooperation additive comprising:
(I) oligosaccharide composition；And/or
(II) carboxylate；With
(III) proteinase complex；(III) proteinase complex is by cultivating the packet dredging continuous shape hyphomycete and obtaining
Proteinase complex containing effective proteinase mixture.Fish special feed cooperation additive helps to improve the micro- life of fish enteron aisle
Object flora, maintains gastrointestinal health and/or reduction pair at the intestinal absorption for promoting the regeneration of fish intestinal villus, improving purpose nutriment
The demand of antibiotic reduces the abuse of antibiotic in breeding process, can also significantly reduce feedstuff-meat ratio when fish culture, shortens and supports
The period is grown, in addition, above-mentioned proteinase complex helps to strengthen the immune system of fish, can obviously reduce and caused by germ contamination
Fish morbidity and mortality, be a kind of efficient fish special feed cooperation additive.
(I) oligosaccharide composition is distributed with following glycosidic bond: there are 8mol% α -1,6 glycosidic bond and β -2 8mol%,
1 glycosidic bond and 8mol% β -1,3 glycosidic bonds, and wherein the degree of polymerization of 8 dry weight % oligosaccharide compositions is 2.
(I) oligosaccharide composition includes: isomaltose-oligosaccharides and isomaltose-inulobiose-oligosaccharides and inulin two
The pool-oligosaccharides and inulobiose-isomaltose-oligosaccharides.Oligosaccharide composition is applied in fish feed additive special or feed
Fish is fed with beneficial health benefit, helps to reduce the fish death rate, improves intestinal microflora, improves nutrients
Matter absorbs, maintains gastrointestinal health and/or reduces the demand to antibiotic, reduces the abuse of antibiotic in breeding process.
(II) carboxylate includes: the lactoyl carboxylate according to formula 1,
Formula 1:R1-COO-[-CH(CH3)-COO]2-R2With its Ba salt, according to the glycollyl carboxylate of formula 2,
Formula 2:R1-COO-[-CH2-COO]3-R2With Ca salt, in above formula, R1For chlorphenyl, the R of formula 12For C2Alkyl
Chain, the R of formula 22For C4Alkyl chain；Above-mentioned carboxylate products application can significantly increase in fish special feed or feed addictive
The antibacterial activity of strong fish, has the function of beneficial in the prevention and treatment of fish enteric infection, and helps to mention
The growth rate of high fish reduces feedstuff-meat ratio and culture-cycle, increases economic benefit.
(III) proteinase complex is to be mixed by what the thin continuous shape hyphomycete of culture obtained comprising effective protease
The proteinase complex of object, the ratio of effective proteinase mixture account for active the 76% of the proteinase complex, institute
In the protease for stating effective proteinase mixture, a kind of protease has 7.4 isoelectric point, and another protease has 8.3
Isoelectric point；Proteinase complex peomotes fish such as fish intestinal villus and regenerates, and therefore dramatically improves purpose battalion
The intestinal absorption of substance is supported, so that feedstuff-meat ratio when significantly reducing fish culture, shortens the culture-cycle, in addition, above-mentioned protease is multiple
Closing object helps to strengthen the immune system of fish, can obviously reduce the disease incidence of the fish as caused by germ contamination, especially extremely
Die rate.
The protease that isoelectric point is 7.4 in (III) proteinase complex has 82000 Anson units/mg protein
Specific activity.
The protease that isoelectric point is 8.3 in (III) proteinase complex has 20000 Anson units/mg protein
Specific activity.
(III) proteinase complex is used in fish special feed as the supplement of feed addictive, the feed
Additive is mixed with fodder compound in powder form.
In order to obtain (III) proteinase complex, cultivate and dredge continuous shape hyphomycete bacterial strain and filtering fermentating liquid, then set about into
Row extracts proteinase complex by ultrafiltration and isoelectric focusing, and finally proteinase complex obtained is lyophilized and crushes, tool
Body includes:
Fermentation: the single colonie scribing line separation of transparent circle, bacterial strain switching after purification are selected from primary dcreening operation culture medium flat plate
Into the fermentation shake flask for filling enzymatic production culture medium, in filtrated air/culture volume/minute ventilation of 0.3 volume,
And ferment at constant temperature 4d under the conditions of enzymatic production medium pH is 7.2, temperature is 40 DEG C, revolving speed is 150r/min, it is taken after centrifugation
Supernatant；
Ultrafiltration: ultrafiltration is carried out by means of the film with 50kDa cutoff threshold, and ultrafiltration carries out 3 hours；
Isoelectric focusing: controlling solution ph respectively is 7.3 and 8.2 progress isoelectric focusing, i.e., what acquisition occupied the majority has 7.4
The protease of left and right isoelectric point (pI), and with less ratio, with 8.3 or so isoelectric points (pI) protease；After freeze-drying crushes
Up to (III) proteinase complex.
The enzymatic production culture medium includes the substance of following weight content: 180 mesh Corncob Xylans 0.18%, threose
Alcohol 0.012%, tryptone 1.8%, yeast extract 0.3%, sodium nitrate 0.1%, KH2PO40.5%, K2HPO40.15%,
MgSO40.03%, natural pH；The carbon source of particular particle size facilitates the output of high enzyme activity protease in enzymatic production culture medium, closes
The carbon source of suitable partial size can make the procreation of microorganism own growth form balance with the dissolved oxygen amount in shaking flask, also can avoid carbon source particle mistake
Generation that is thin and simultaneously influencing Metabolite in turn with other nutriments formation biggish suspension of viscosity, reduces producing enzyme water
It is flat；In addition, adding the production of the Corncob Xylan and threitol of special proportion to high enzyme activity protease in enzymatic production culture medium
Induction stimulation has been provided, can greatly accelerate point for dredging the highly active protein enzyme that continuous shape hyphomycete bacterial strain is 7.4 to isoelectric point
It secretes and is precipitated, to significantly improve the protease that isoelectric point is 7.4 on the basis of maintaining the content of whole proteinase complex
Accounting, on the whole improve proteinase complex activity.
Comparative example 1:
Comparative example 1 and embodiment 1 are essentially identical, the difference is that: do not contain threitol in enzymatic production culture medium.
Embodiment 2:
One kind (I) oligosaccharide composition is distributed with following glycosidic bond: having 15mol% α -1,6 glycosidic bond and 12mol%
β -2,1 glycosidic bond, and wherein the degree of polymerization of 10 dry weight % oligosaccharide compositions is 6.
(I) oligosaccharide composition includes: isomaltose-galacto-oligosaccharide, lactose-inulobiose-oligosaccharides and inulin two
Sugar-isomaltose-oligosaccharides.Oligosaccharide composition is applied in fish feed additive special or feed feeds fish with beneficial
Health benefit, facilitate reduce the fish death rate, improve intestinal microflora, improve absorption of nutrient ingredients, maintain stomach and intestine
The demand of road health and/or reduction to antibiotic, reduces the abuse of antibiotic in breeding process.
A kind of (II) carboxylate are as follows: according to the lactoyl carboxylate of formula 1,
Formula 1:R1-COO-[-CH(CH3)-COO]2-R2With its Na salt, according to the glycollyl carboxylate of formula 2,
Formula 2:R1-COO-[-CH2-COO]4-R2With its NH4Salt；In above formula, the R of formula 11For chlorphenyl, the R of formula 12For
C3Alkenylene chain, the R of formula 21For bromophenyl, the R of formula 22For C4Alkyl chain；Above-mentioned carboxylate products application in fish special feed or
In person's feed addictive, the antibacterial activity of fish can be significantly increased, is had in the prevention and treatment of fish enteric infection
Beneficial effect, and the growth rate of fish is helped to improve, feedstuff-meat ratio and culture-cycle are reduced, economic benefit is increased.
A kind of (III) includes the proteinase complex of effective proteinase mixture, is obtained by cultivating thin continuous shape hyphomycete
, the ratio of effective proteinase mixture accounts for active the 82% of the proteinase complex, and effective protease is mixed
In the protease for closing object, a kind of protease has 7.4 isoelectric point, and another protease has 8.3 isoelectric point；Protease
Compound peomotes fish such as fish intestinal villus and regenerates, and therefore dramatically improves the intestinal absorption of purpose nutriment,
To significantly reduce feedstuff-meat ratio when fish culture, shorten the culture-cycle, in addition, above-mentioned proteinase complex helps to strengthen
The immune system of fish can obviously reduce the disease incidence of the fish as caused by germ contamination, the especially death rate.
The protease that isoelectric point is 7.4 or so in (III) proteinase complex has 85000 Anson units/mg egg
The specific activity of white matter.
The protease that isoelectric point is 8.3 or so in (III) proteinase complex has 22000 Anson units/mg egg
The specific activity of white matter.
(III) proteinase complex is used in fish special feed as the supplement of feed addictive, the feed
Additive is mixed with fodder compound in powder form.
In order to obtain (III) proteinase complex, cultivate and dredge continuous shape hyphomycete bacterial strain and filtering fermentating liquid, then set about into
Row extracts proteinase complex by ultrafiltration and isoelectric focusing, and finally proteinase complex obtained is lyophilized and crushes, tool
Body includes:
1 fermentation: the single colonie of transparent circle, scribing line separation are selected from primary dcreening operation culture medium flat plate；Bacterial strain after purification turns
It is connected in the fermentation shake flask for filling enzymatic production culture medium, in filtrated air/culture volume/minute ventilation of 0.3 volume,
And ferment at constant temperature 6d under the conditions of enzymatic production medium pH is 7.5, temperature is 45 DEG C, revolving speed is 200r/min, it is taken after centrifugation
Supernatant；
2 ultrafiltration: ultrafiltration is carried out by means of the film with 50kDa cutoff threshold, and ultrafiltration carries out 10 hours；
3 isoelectric focusing: controlling solution ph respectively is 7.5 and 8.4 progress isoelectric focusing, i.e., what acquisition occupied the majority has
The protease of 7.4 or so isoelectric points (pI), and with less ratio, the protease with 8.3 or so isoelectric points (pI)；Freeze-drying crushes
Afterwards up to (III) proteinase complex.
Enzymatic production culture medium includes the substance of following weight content: 240 mesh Corncob Xylans 0.2%, threitol
0.014%, tryptone 3%, yeast extract 0.5%, sodium nitrate 0.2%, KH2PO40.6%, K2HPO40.2%,
MgSO40.05%, natural pH；The carbon source of particular particle size facilitates the output of high enzyme activity protease in enzymatic production culture medium, closes
The carbon source of suitable partial size can make the procreation of microorganism own growth form balance with the dissolved oxygen amount in shaking flask, also can avoid carbon source particle mistake
Generation that is thin and simultaneously influencing Metabolite in turn with other nutriments formation biggish suspension of viscosity, reduces producing enzyme water
It is flat；In addition, adding the production of the Corncob Xylan and threitol of special proportion to high enzyme activity protease in enzymatic production culture medium
Induction stimulation has been provided, can greatly accelerate the highly active protein enzyme for dredging that continuous shape hyphomycete bacterial strain is 7.4 or so to isoelectric point
Secretion and precipitation, so that significantly improving isoelectric point on the basis of maintaining the content of whole proteinase complex is 7.4 or so
Protease accounting, on the whole improve proteinase complex activity.
A kind of fish special feed cooperation additive comprising above-mentioned:
(I) oligosaccharide composition, and (II) carboxylate, and (III) proteinase complex；(III) proteinase complex is
By cultivating the proteinase complex comprising effective proteinase mixture dredging continuous shape hyphomycete and obtaining.Fish special feed is matched
Closing additive facilitates the intestines for improving fish intestinal microflora, promoting the regeneration of fish intestinal villus, improving purpose nutriment
The demand to antibiotic is absorbed, maintains gastrointestinal health and/or reduced, the abuse of antibiotic in breeding process is reduced, can also show
Feedstuff-meat ratio when reducing fish culture is write, the culture-cycle is shortened, in addition, above-mentioned proteinase complex helps to strengthen exempting from for fish
Epidemic disease system can obviously reduce the morbidity and mortality of the fish as caused by germ contamination, be a kind of dedicated feeding of efficient fish
Material cooperation additive.
Comparative example 2:
Comparative example 2 and embodiment 2 are essentially identical, the difference is that: it is imbeautiful containing 240 mesh in enzymatic production culture medium
Rice core xylan.
Embodiment 3:
A kind of fish special feed cooperation additive comprising:
(I) oligosaccharide composition；(II) carboxylate；(III) proteinase complex；(III) proteinase complex is
By cultivating the proteinase complex comprising effective proteinase mixture dredging continuous shape hyphomycete and obtaining.Fish special feed is matched
Closing additive facilitates the intestines for improving fish intestinal microflora, promoting the regeneration of fish intestinal villus, improving purpose nutriment
The demand to antibiotic is absorbed, maintains gastrointestinal health and/or reduced, the abuse of antibiotic in breeding process is reduced, can also show
Feedstuff-meat ratio when reducing fish culture is write, the culture-cycle is shortened, in addition, above-mentioned proteinase complex helps to strengthen exempting from for fish
Epidemic disease system can obviously reduce the morbidity and mortality of the fish as caused by germ contamination, be a kind of dedicated feeding of efficient fish
Material cooperation additive.
(I) oligosaccharide composition is distributed with following glycosidic bond: having 20mol% α -1,6 glycosidic bonds and 20mol%
β -2,1 glycosidic bond and 20mol% β -1,3 glycosidic bonds, and wherein the degree of polymerization of 13 dry weight % oligosaccharide compositions is 4.
(I) oligosaccharide composition includes: isomaltose-oligosaccharides, isomaltose-inulobiose-oligosaccharides, two pool of inulin-
Oligosaccharides, two pools of chondroitin-oligosaccharides, two pools of hyalomitome-oligosaccharides, two pools of hyalomitome-isomaltose-oligosaccharides, two pool of chondroitin-different wheat
Bud sugar-oligosaccharides, galacto-oligosaccharide, lactose-two pools of hyalomitome-oligosaccharides, isomaltose-galacto-oligosaccharide, lactose-inulobiose-widow
Sugar and inulobiose-isomaltose-oligosaccharides.Oligosaccharide composition is applied in fish feed additive special or feed to feed
Fish has beneficial health benefit, helps to reduce the fish death rate, improves intestinal microflora, improves nutriment suction
The demand to antibiotic is received, maintains gastrointestinal health and/or reduced, the abuse of antibiotic in breeding process is reduced.
(II) carboxylate is selected from: according to the lactoyl carboxylate of formula 1,
Formula 1:R1-COO-[-CH(CH3)-COO]3-R2With its Mn (II) salt, according to the glycollyl carboxylate of formula 2, formula 2:R1-
COO-[-CH2-COO]6-R2With its Co (II) salt, in above formula, the R of formula 1 and formula 21For bromophenyl, the R of formula 12For C5Alkane
Base, the R of formula 22For C3Alkenylene chain；Above-mentioned carboxylate products application can significantly increase in fish special feed or feed addictive
The antibacterial activity of strong fish, has the function of beneficial in the prevention and treatment of fish enteric infection, and helps to mention
The growth rate of high fish reduces feedstuff-meat ratio and culture-cycle, increases economic benefit.
(III) proteinase complex is to be mixed by what the thin continuous shape hyphomycete of culture obtained comprising effective protease
The proteinase complex of object, the ratio of effective proteinase mixture account for active the 85% of the proteinase complex, institute
In the protease for stating effective proteinase mixture, a kind of protease has 7.4 isoelectric point, and another protease has 8.3
Isoelectric point；Proteinase complex peomotes fish such as fish intestinal villus and regenerates, and therefore dramatically improves purpose battalion
The intestinal absorption of substance is supported, so that feedstuff-meat ratio when significantly reducing fish culture, shortens the culture-cycle, in addition, above-mentioned protease is multiple
Closing object helps to strengthen the immune system of fish, can obviously reduce the disease incidence of the fish as caused by germ contamination, especially extremely
Die rate.
The protease that isoelectric point is 7.4 or so in (III) proteinase complex has 88000 Anson units/mg egg
The specific activity of white matter.
The protease that isoelectric point is 8.3 or so in (III) proteinase complex has 24000 Anson units/mg egg
The specific activity of white matter.
(III) proteinase complex is used in fish special feed as the supplement of feed addictive, the feed
Additive is mixed with fodder compound in powder form.
In order to obtain (III) proteinase complex, cultivate and dredge continuous shape hyphomycete bacterial strain and filtering fermentating liquid, then set about into
Row extracts proteinase complex by ultrafiltration and isoelectric focusing, and finally proteinase complex obtained is lyophilized and crushes, tool
Body includes:
1 fermentation: single colonie (the decomposable culture of the produced protease of bacterial strain of transparent circle is selected from primary dcreening operation culture medium flat plate
Substrate protein in base, therefore transparent circle phenomenon is presented in periphery of bacterial colonies), scribing line separation；Bacterial strain after purification, which is forwarded to, fills fermentation
In the fermentation shake flask of culture medium, produced in filtrated air/culture volume/minute ventilation of 0.3 volume, and in fermentation
Ferment at constant temperature 5d under the conditions of enzyme medium pH is 7.4, temperature is 42 DEG C, revolving speed is 180r/min, takes supernatant after centrifugation；
2 ultrafiltration: ultrafiltration is carried out by means of the film with 50kDa cutoff threshold, and ultrafiltration carries out 5 hours；
3 isoelectric focusing: controlling solution ph respectively is 7.4 and 8.3 progress isoelectric focusing, i.e., what acquisition occupied the majority has
The protease of 7.4 or so isoelectric points (pI), and with less ratio, the protease with 8.3 or so isoelectric points (pI)；Freeze-drying crushes
Afterwards up to (III) proteinase complex.
Enzymatic production culture medium includes the substance of following weight content: 200 mesh Corncob Xylans 0.18%, threitol
0.012%, tryptone 2%, yeast extract 0.4%, sodium nitrate 0.15%, KH2PO40.55%, K2HPO40.18%,
MgSO40.04%, natural pH；The carbon source of particular particle size facilitates the output of high enzyme activity protease in enzymatic production culture medium, closes
The carbon source of suitable partial size can make the procreation of microorganism own growth form balance with the dissolved oxygen amount in shaking flask, also can avoid carbon source particle mistake
Generation that is thin and simultaneously influencing Metabolite in turn with other nutriments formation biggish suspension of viscosity, reduces producing enzyme water
It is flat；In addition, adding the production of the Corncob Xylan and threitol of special proportion to high enzyme activity protease in enzymatic production culture medium
Induction stimulation has been provided, can greatly accelerate the highly active protein enzyme for dredging that continuous shape hyphomycete bacterial strain is 7.4 or so to isoelectric point
Secretion and precipitation, so that significantly improving isoelectric point on the basis of maintaining the content of whole proteinase complex is 7.4 or so
Protease accounting, on the whole improve proteinase complex activity.
Comparative example 3:
Comparative example 3 and embodiment 3 are essentially identical, the difference is that: 200 mesh corns are not contained in enzymatic production culture medium
Core xylan and threitol.
Experimental example:
1, detecting one of (III) proteinase complex isoelectric point in Examples 1 to 3 and comparative example 1~3 respectively is 7.4
The protease that the protease of left and right and another isoelectric point are 8.3 or so each account for than and specific activity, statistics it is as shown in table 1.
The accounting and specific activity of 1~3 protease of 1 Examples 1 to 3 of table and comparative example
As shown in Table 1, in the preferred embodiment of the present invention 1~3, pI be 7.4 or so highly active protein enzyme accounting compared with
Height, and its activity is relatively stable, largely improves the whole enzymatic activity of (III) proteinase complex.
2, a kind of fish special feed adds containing basal feed and the cooperation of fish special feed in the fish special feed
Add agent, the basal feed includes: the fat of the protein of 32wt%, 22wt%；Nucleotide, the 4wt% of 0.08wt% can disappear
Total effective calcium, total available phosphorus of 0.35wt% and the sodium of 0.3wt% of the sulfur-containing amino acid of change, 0.4wt%.
The fish special feed cooperation additive is respectively selected from Examples 1 to 3 and comparative example 1~3.
Above-mentioned 6 kinds of fish special feeds are fed respectively and give cultivation fish, other factors are completely the same, feed 4 months
Afterwards, average body length and average weight and the death rate, the statistical result for measuring fish respectively are as shown in table 2.
The cultivation situation of 2 fish of table counts
| Measurement item | Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example 1 | Comparative example 2 | Comparative example 3 |
| Average body is long (cm) | 40.5 | 41.6 | 43.0 | 35.5 | 34.2 | 33.8 |
| Average weight (g) | 855.1 | 860.9 | 905.5 | 705.8 | 710.6 | 700.2 |
| The death rate (%) | 0.5% | 1.0% | 0.5% | 3.5% | 4.0% | 4.5% |
As shown in Table 2, average body length, average weight and the survival rate of fish are significantly better than comparative example, illustrate the present invention
In fish special feed cooperation additive can effectively improve the growth rate of fish, reduce feedstuff-meat ratio and culture-cycle, be
A kind of fish special feed cooperation additive efficiently, economic.
The prior art of routine techniques dawn known to those skilled in the art in above-described embodiment, therefore herein no longer in detail
It repeats.
The above embodiments are only used to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this field
Member can also make a variety of changes and modification without departing from the spirit and scope of the present invention.Therefore, all equivalent
Technical solution also belong to scope of the invention, scope of patent protection of the invention should be defined by the claims.
Claims (10)
1. fish special feed cooperates additive, characterized by comprising:
(I) oligosaccharide composition；And/or
(II) carboxylate；With
(III) proteinase complex；
(III) proteinase complex is to be obtained by the thin continuous shape hyphomycete of culture comprising effective proteinase mixture
Proteinase complex.
2. fish special feed according to claim 1 cooperates additive, it is characterised in that: (I) oligosaccharide composition
It is distributed: having with following glycosidic bond
No less than 8mol% α -1,6 glycosidic bonds, and
No less than 8mol% β -2,1 glycosidic bond, and/or
No less than 8mol% β -1,3 glycosidic bonds, and
The degree of polymerization of at least 8 dry weight % oligosaccharide compositions is 2~6 in (I) oligosaccharide composition.
3. fish special feed according to claim 1 or 2 cooperates additive, it is characterised in that: (I) the oligosaccharides combination
Object include: isomaltose-oligosaccharides, isomaltose-inulobiose-oligosaccharides, two pools of inulin-oligosaccharides, two pools of chondroitin-oligosaccharides, thoroughly
Bright two pools of matter-oligosaccharides, two pools of hyalomitome-isomaltose-oligosaccharides, two pools of chondroitin-isomaltose-oligosaccharides, galacto-oligosaccharide, cream
Sugar-two pools of hyalomitome-oligosaccharides, isomaltose-galacto-oligosaccharide, lactose-inulobiose-oligosaccharides, inulobiose-isomaltose-widow
Sugar.
4. fish special feed according to claim 1 cooperates additive, it is characterised in that: (II) the carboxylate choosing
From:
According to the lactoyl carboxylate of formula 1,
Formula 1:R1-COO-[-CH(CH3)-COO]n-R2
Or/and its Ba, Be, Mn (II), Ca, Na, NH4Or Co (II) salt,
According to the glycollyl carboxylate of formula 2,
Formula 2:R1-COO-[-CH2-COO]n-R2
Or/and its Ba, Be, Mn (II), Ca, Na, NH4Or Co (II) salt,
In above formula, R1Selected from chlorphenyl or bromophenyl, R2Selected from C2~C10Alkyl or alkenylene chain, and n indicates that numerical value is
1~10 integer.
5. fish special feed according to claim 1 cooperates additive, it is characterised in that: (III) protease is compound
The ratio of effective proteinase mixture is more than active the 75% of the proteinase complex in object, effective protease mixing
In the protease of object, a kind of protease has 7.4 or so isoelectric point, and another protease has 8.3 or so isoelectric point.
6. fish special feed according to claim 5 cooperates additive, it is characterised in that: (III) protease is compound
The protease that isoelectric point is 7.4 or so in object has 82000~90000 Anson units/mg protein specific activity, described (III)
The protease that isoelectric point is 8.3 or so in proteinase complex is living with 20000~25000 Anson units/mg protein ratio
Property.
7. fish special feed according to claim 5 or 6 cooperates additive, it is characterised in that: (III) protease
Compound as feed addictive supplement in fish special feed, the feed addictive with powder, liquid or its
He is any to be suitble to the form mixed with fodder compound to exist.
8. according to claim 1,2, the described in any item fish special feeds cooperation additives of 4-6, it is characterised in that: described
(III) preparation process of proteinase complex includes:
Fermentation: the thin continuous shape hyphomycete bacterial strain single colonie scribing line separation of transparent circle is selected from primary dcreening operation culture medium flat plate；Purifying
Bacterial strain afterwards, which is forwarded in the fermentation shake flask for filling enzymatic production culture medium, to ferment, and supernatant is taken after centrifugation；
Ultrafiltration: ultrafiltration is carried out by means of the film with 50kDa cutoff threshold；
Isoelectric focusing: controlling solution ph respectively is 7.4 ± 0.1 and 8.3 ± 0.1 progress isoelectric focusing, that is, is obtained left with 7.4
The protease of right isoelectric point (pI), and the protease with 8.3 or so isoelectric points (pI)；Up to (III) protease after freeze-drying crushing
Compound.
9. fish special feed according to claim 8 cooperates additive, it is characterised in that: the enzymatic production culture medium
Substance including following weight content: 180~240 mesh Corncob Xylans 0.18~0.2%, threitol 0.012~
0.014%, tryptone 1.8~3%, yeast extract 0.3~0.5%, sodium nitrate 0.1~0.2%, KH2PO40.5~0.6%,
K2HPO40.15~0.2%, MgSO40.03~0.05%, natural pH.
10. the fish special feed comprising the described in any item fish special feed cooperation additives of claim 1-9, feature
It is containing basal feed and such as the described in any item fish special feed cooperation additives of claim 1-9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811374998.6A CN109757610B (en) | 2018-11-19 | 2018-11-19 | Special feed additive for brown croakers |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811374998.6A CN109757610B (en) | 2018-11-19 | 2018-11-19 | Special feed additive for brown croakers |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109757610A true CN109757610A (en) | 2019-05-17 |
| CN109757610B CN109757610B (en) | 2022-05-06 |
Family
ID=66449557
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811374998.6A Active CN109757610B (en) | 2018-11-19 | 2018-11-19 | Special feed additive for brown croakers |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109757610B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2200344A1 (en) * | 1994-10-13 | 1996-04-25 | Anne-Cathrine Samuelsson | Animal feed of higher nutritive value, method for production thereof and use of a polyethylene glycol compound |
| CN101326958A (en) * | 2008-07-22 | 2008-12-24 | 浙江大学 | Peptide feedstuff additive for regulating immunity function of sea water fish |
| CN102362635A (en) * | 2011-10-18 | 2012-02-29 | 珠海市世海饲料有限公司 | High-quality compound feed with low nitrogen and phosphorus emission for Lateolabrax |
-
2018
- 2018-11-19 CN CN201811374998.6A patent/CN109757610B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2200344A1 (en) * | 1994-10-13 | 1996-04-25 | Anne-Cathrine Samuelsson | Animal feed of higher nutritive value, method for production thereof and use of a polyethylene glycol compound |
| CN101326958A (en) * | 2008-07-22 | 2008-12-24 | 浙江大学 | Peptide feedstuff additive for regulating immunity function of sea water fish |
| CN102362635A (en) * | 2011-10-18 | 2012-02-29 | 珠海市世海饲料有限公司 | High-quality compound feed with low nitrogen and phosphorus emission for Lateolabrax |
Non-Patent Citations (1)
| Title |
|---|
| 李旺等: "饲料中菌酶协同作用的研究与应用进展", 《动物营养学报》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109757610B (en) | 2022-05-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104799124B (en) | A kind of colored clam mixed feed | |
| CN104431621B (en) | A kind of aquatic products compound premix and preparation method thereof | |
| CN103349157B (en) | Compound feed for female parent fishes of sturgeons | |
| CN103704530B (en) | A kind of loach seedling special compound feed | |
| CN103355491A (en) | Alga immunopotentiation compound feed for litopenaeus vannamei | |
| CN110731417B (en) | Microbial fermentation feed for crayfish and preparation method thereof | |
| CN106343257A (en) | Feed capable of promoting molt of crayfish and preparation method thereof | |
| CN106260788A (en) | A kind of Micropterus salmonoides premix material | |
| CN108271938A (en) | A kind of functional seaweed fodder additive improving prevention of sow constipation | |
| CN113080339A (en) | Special compound feed for industrial aquaculture of penaeus monodon | |
| CN107348229A (en) | A kind of catfish feed eating belt agent and preparation method thereof | |
| KR20110099872A (en) | A feed additive for raising fishes and a method of manufacturing for a feed for raising fishes | |
| CN109907163A (en) | Feed addictive of layer chicken and method containing calcium citrate malate are prepared using shrimp and crab shells decalcification | |
| CN107251858B (en) | Method for breeding epinephelus coioides | |
| CN106819536A (en) | Green nourishing feed for producing just raw egg laying hen | |
| CN102224882A (en) | Preparation method for feed additive with antioxidation function | |
| CN105010757B (en) | Application of the polymalic acid calcium in fodder acidulant is prepared | |
| CN109757610A (en) | Fish special feed cooperates additive | |
| CN111418712A (en) | Feed additive for improving survival rate of weever and preparation method thereof | |
| CN106819617A (en) | A kind of rheum officinale fish meal of growth promotion diseases prevention and its application | |
| CN113558150A (en) | Feed nutrient containing polyglutamic acid and application thereof | |
| CN111084297A (en) | Feed additive for preventing and treating enteritis of pikes | |
| CN109430547A (en) | Small water turtle children's tortoise puffing granule compound feed and preparation method thereof | |
| CN103005178A (en) | Prawn nutrition intensifier | |
| CN113519717B (en) | Composite feed additive and feed for laying hens in growing period |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |