NZ708401B2 - Use of an enzymatic complex in the feeding of livestock animals - Google Patents
Use of an enzymatic complex in the feeding of livestock animals Download PDFInfo
- Publication number
- NZ708401B2 NZ708401B2 NZ708401A NZ70840113A NZ708401B2 NZ 708401 B2 NZ708401 B2 NZ 708401B2 NZ 708401 A NZ708401 A NZ 708401A NZ 70840113 A NZ70840113 A NZ 70840113A NZ 708401 B2 NZ708401 B2 NZ 708401B2
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- New Zealand
- Prior art keywords
- complex
- feed
- enzymatic
- enzymatic complex
- around
- Prior art date
Links
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- 241001465754 Metazoa Species 0.000 title claims abstract description 19
- 244000144972 livestock Species 0.000 title abstract description 5
- 239000000203 mixture Substances 0.000 claims abstract description 42
- 108091005771 Peptidases Proteins 0.000 claims abstract description 32
- 239000004365 Protease Substances 0.000 claims abstract description 30
- 230000000694 effects Effects 0.000 claims abstract description 28
- 102000035443 Peptidases Human genes 0.000 claims abstract description 20
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 19
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 19
- 241000187438 Streptomyces fradiae Species 0.000 claims abstract description 13
- 238000004519 manufacturing process Methods 0.000 claims abstract description 6
- 102000033147 ERVK-25 Human genes 0.000 claims description 12
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/189—Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
Abstract
The invention relates to the use of an enzymatic complex comprising a mixture of proteases, which is obtained by culturing a strain of Streptomyces fradiae, for supplementing the feeding of livestock animals, in which one of the proteases has an isoelectric point of about 7.0 and a specific activity of around 150,000 Anson units/mg of protein and another of the proteases has an isoelectric point of about 8.0 and a specific activity of around 38,000 Anson units/mg of protein, wherein greater than 80% of the activity of said enzymatic complex is due to these two types of proteases. The invention also relates to feed compositions for livestock animals comprising said enzymatic complex and also to the method for producing this complex. of around 150,000 Anson units/mg of protein and another of the proteases has an isoelectric point of about 8.0 and a specific activity of around 38,000 Anson units/mg of protein, wherein greater than 80% of the activity of said enzymatic complex is due to these two types of proteases. The invention also relates to feed compositions for livestock animals comprising said enzymatic complex and also to the method for producing this complex.
Description
USE OF AN ENZYMATIC COMPLEX IN THE FEEDING OF LIVESTOCK ANIMALS
The technical field of the present ion is that of
animal nutrition.
It is known that the Streptomyces fradiae strain is able
to produce at least five types of protease known by Ia, Ib,
II, III, IV and two peptidases (patent GB-1133579)
In their patent application WO 87/07905, the applicants
described a process to obtain a lytic x composed
exclusively of type II, III and IV proteases with a
predominance of type I I se. This complex is obtained
from a Streptomyces fradiae WAKSMAN 3535 strain by
fermentation, extraction of the complex by filtration, then
ultrafiltration on a ne having a cut-off level
corresponding to a molecular weight of between 2,000 and
12,000, and finally by atomization. The complex thus obtained
in powder form has a titer of at least 2,000 Anson units/mg
of protein.
In this application, the type II protease was
characterized by a molecular weight approaching 18 kDa and an
isoelectric point approaching 9.0.
This complex, used in animal foodstuffs, enables the
animals to easily assimilate feed with high crude protein
t thereby promoting their growth. Additionally, this
complex, used for ing sows, promotes increased birth
weights as well as a reduction in food consumption and an
overall improvement in health.
r, this process is not able to effectively
eliminate further contaminating ducts.
In patent application WO 97/37681, improvements are made
enabling the use of a product with a titer of at least 4,000
Anson units/mg of protein for therapeutic ations. Thus,
the predominant proteinase has a specific activity of over
100,000 Anson units/mg of protein with a molecular weight of
between 30 and 36 kDa and an isoelectric point approaching
7.0.
Subsequent analyses showed that the molecular weight of
the protease was of 18-20 kDa.
The tterapeutic applications described essentially
consist of a posi:ive action in the reduction of the
secretion of mucus cnaracteristic of certain illnesses, in an
improvement in intestinal absorption and blood flow or e"se
in the figrt against infectious diseases. Additionally, the
compositions are described as able to facilitate the
regeneration of atrophied or damaged villi in the intestinal
lining.
"n view of the considerable therapeutic ties of the
enzyma:ic complex e xtraCCed from Streptomyces fradiae
cultures, it is important to use an enzymatic complex more
completely cleared of biological impurities that might hinder
its ac:ivity.
The aim of the t invention is thus to improve the
nsly described. enzymatic x namely in order to
enhance breeding conditions and animal health in factory
farms.
The invention thus relates to the utilization of an
enzymatic complex comprising a mixture of proteases obtained
by culturing a Streptomyces fradiae strain to supplement farn
animals feed, characterized in that among this ndxture o;
proteases one of them has an isoelectric point of around 7.0
and another has an isoelectric point 0: around 8.3.
Said complex predominantly contains these two types of
proteases, in o:her words a proportion of these two ses
exceeding 80% of the activity of said mixture.
According' to one characteristic of the invention, the
protease whose isoelectric point is of around 7.0 has a
specific activity 0: around 150,000 Anson units/mg of
pro:ein.
ing to one characteristic of the invention, the
pro:ease whose isoelectric point is of around 8.0 has a
specific activity of around 38,000 Ansor mg of protein.
According to another characteristic of the invention, the
enzymatic complex is used as a feed supplement in ,
liquid or any other form suited to its deture with feed
compositions to improve the general condition o_ factory farm
animals.
According to another charaCteristic of the invention, the
enzymatic complex in powder form is dry mixed with a feed
composition in a rotating drum anti; homogenized.
ing to aqother cqaraCteristic of tre invention, the
enzymatic complex in liquid form is pulveri ed in a f“uidi ed
bed onto a feed conpositioq, and is then grarulated.
According to another Ciaracteristic of the invention, the
enzymatic complex is used in poultry feed.
According to r oiaracteriStic of tre invention, the
enzymatic x is used in pig feed.
ing to aiother cqaracteriStic of tre invention, the
enzymatic complex is used in fish feed.
The invention also relates to a feed composition for farm
animals comprising the enzymatic complex ing to the
ion and possibly including excipients or vehicles for
nutriments, or other additives.
r1"he invention lastly relates to a manufacturing process
for the enzymatic complex according to the invention,
characterized in that a Streptomyces fradiae strain is
cultured, in that the fermentation broth is filtered, in that
the enzymatic complex is thereafter extraCted by
ultrafiitration and ion exchange chromatography followed by
an isoeiectric focusing and finally, the x thus
obtained is ‘yophiii ed.
This invention provides a feed ment whose
characteristics are strictly defined.
The invention firstly enables to adapt precisely the
doses to be administered into the feed compositions so as to
obtain thc r guirod thorapcutic effects.
The invention also has the advantage 0: improving the
health of animals in the y farms.
in addition, the invention also brings ar improvement to
the general condition of the animals in the factory farms
enhancing the profitability 0‘ such facilities.
The invention further enables a1 improved animal growth
combined with a reduction in food ption.
Other teristics, ulars and advantages 0: the
invention will become more apparent from, the additional
description given hereafter of di ,— ferent embodiments given by
way 0: e and with respect to the figures illustrating
inteStinal villi.
it is known that the enzyme mixture commercialized under
the trademark Panstimase® (PANcreatic ST"Mu"ating diastASE)
is an enzymatic x ed by fermertation of
Streptomyces fradiae. This complex is described as
essentially containing three proteases, n of wiich
specifically act on tial proteins gen, elaStin,
and keratin), but also on certain toxins of proteic nature
ed by choleraic vibrio or certain pathogenic
Escherichia coli strains.
in the domain of animal husbandry, it is important to
have a product that is technically defined, perfeCtly
conStant and whose ties are thus clearly defined.
So as to obtain the enzymatic complex according to the
invention a Streptomyces fradiae strain is cultured and the
resulting fermentation broth is filtered. Thereafter, the
enzymatic complex is extracted and terized by ultra—
filtration, ion exchange chromatography and isoelectnic
focusing. Lastly, the resulting complex is lfi ed.
For the Streptomyces fradiae culture, a strain. of the
type WAKSMAN 353 for example is used, able to be genetically
improved. to namely obtain an increase in the proteolytic
activity or the suppression of any secretion of antibiotics.
The culture takes place in a large scale fermenter ir a
suitable growing medium. This growing medium may, for
example, be based on soybean meal: 15 g/l; glucose: 30 g/l;
bipotassic phosphate: 1 g/l; calcium carbonate: 10 g/l.
The reaction takes place at a pH 0: between 7.0 and 7.5,
at a fermentation temperature 0: around 28 0C with an
on of 0.3 s of steri e air for one volume of
mediJm per minute.
Once is culture is completed, the mycelium is eliminated
by filtration of the fermentation broth using a clarifying
agent.
The extraction of the enzymatic complex required the
implementation of several techniques including ultra—
tion, ion exchange chromatography and isoelectric
focusing.
Ultra—filtration
The ul:ra—filtration of she fil:ra:e is performed using a
membrane With a CUt‘Om’ “eve of 50 (Da.
After 3h of filtering, the fil':ra:e (71% of the l
volume) has a proteolytic activity reveaj_ed by the gelatin
*i'm assay. This assay consists ir deteriorating a film of
silver gelatin arranged on a solid frame.
ion exchange chromatography assay
A cation exchange chromatography is used made with
colu ns of Cellu fine C—500 carboxymethy: gel (Amicon) o;
suitable dimensiors.
The chromatographic bu” .Eer is a citrate bu ,. "er 10 mM at
pH 5.5 and the elution is performed by a NaCl gradient 0: 0
to 1.0 M. The elution rate s on the size of the
columns, for example, it is of l0 to 15 ml/min for a column
0: 2.5 cm in diameter and 27 on in height.
Isoelectric focusing
2O ical ctric ’oc ‘ng is per-formed using ready—
to—use gels made by Pharmacia for pHs from 3 to 9 on Phast—
system. tion is per:formed using Coomassie blue.
in the case o: preparative isoelectric ng in a
solid medium, the enzyme solution is dialyzed against
distilled water (95 ml) and mixed with 5 ml 0:: 3—9 ampholytes
with 4.7 g o: Ultrodex gel and 0.5 g o: glycine. The gel is
dried for 6 h at 40°C. Migration takes place during the
night, or equiva lent period, at a cor stant power of 3 Vt
Revelation is per:formed us ing Coomassie blue.
The different gel strf-ps are taken, rinsed with 3 ml 0:
water, filtered, and then rinsed with 3 ml of G:
buffer.
The fractions thus obtained are then analyzed to
determine their proteolytic activity and protein content. The
s are then dialyzed against the Tris—HG: buffer 0.3 M
pH 8.0.
In the case o: preparative isoe‘ectric focusing in a
liquid medium, the enzyme solution dia:_yzed against distilled
water is mixed with the ampholytes (3.5 ml). Migration is
performed at a constant power of 8 W during ~ h.
The fraction s amples are ed then dialyzed against
the TriS*HC" buffer 0.3 M pH 8.0.
The experimental conditions indicated above are to be
adapted according to the quantities of product required to be
obtained and to its utilization in rial conditions.
A predominant protease is thereby obtained in the mix:ure
that has an isoelectric point (:EP) 0: around 7.0 and in a
lesser propor:ion a protease whose iiP is 0: around 8.0.
The predominant protease whose "?P is 0“ around 7.0 is
obtained in a volume of 9 ml with a protein tration 0;
0.49 mg/ml. Speci a; precautions for purification may be
required because 0; :he itation of this protease at its
isoelectric point. The solution 0btained has a titer of
around 75,000 Anson inits (A.U)/ml.
This protease is purified after homogenization at a scale
of 4 mg with a specific activity of 150,000 A.U/mg of
protein.
One Anson unit (AU) is defined here as being the quantity
of enzyme which, incubated for 10 minutes at 25 °C and at pH
7.5 in the C
presence o; deratured hemoglobin, releases from
this substrate the equivalent of l. microgranl of tyrosine,
determined by spectro—photometric absorption at 280 nm on the
filtrate that cannot be precipitated with trichloracetic
acid.
rThe protease, whose ifip is O“ around 8.0, is recovered
join:ly with the previous one but in a greater volume, 0: 35
ml for example, and has a protei n concentration of 1.48
3O mg/mi. This solution ras a titer of around 56,000 A.U/ml.
This protease i s ed afte” homogenization at a scale
o: 50 mg with a ic activity 0:5 around 38,000 A.U/mg of
protein.
The previously described. enzymatic complex essentially
contains pro:eases and other proteins. The purifications
performed and ':he ophoresis technique used. prove an
additional purification and the elimination of inating
als that have no proteolytic activity.
The different proteases ed may be better
characterized by determining tha: activities on proteins such
as collagen, <eratin, n or hemoglobin.
Unfortunately, these dete”minat ons are complicated
because of their lack of ivity and specificity.
se m 4st ore be made to several activity assays,
constituting detection methods, and principally the Anson
method.
Activity assays
Anson assay
This assay enables a proteolytic activity to be dosed,
without icity. "t consists in dosing the hydrolysis of
the denatured hemoglobin by spectrophotometric determination
0: the tyrosine contained in the oligopeptides released.
Azocasein assay
This assay consists in determining by spectrophotometry
at 440 nm the hydro“ ysis C
0’ the casein stained by an orange
dye. This assay is not speci fic. After incubation with :he
enzyme, the remaining azocasein is itated using
:richloroacetic acid. irg the optical y reflects
:he presence of the peptides released, and thus the enzymatic
activity.
Elastolxtic activity assay
Elastin Congo—Red assay
This assay consists in spectrophotometrically dosirg at
495 im the stained peptides released by the hydrolysis of the
elas:in stained by a red dye.
A possible problen1 of this dosage lies in :he
insolubility of the substrate and thus its sensitivity
depends on the sample accuracy, :he stirring of :he
tion mediim and the adherence of the substrate :0 the
walls of the incubation vessel.
The assays performed show a lack of linearity between the
activity and quantity of enzyme. There is e”"ectively
linearity between the activity and the incubation time, but
the straight line representing it does not pass through the
origin.
A on: Serence in absorbance can thus be measured between
the incuba: ion times (2O and. 40 minutes). With 10 ul of
complex d 10 times, a variation 0: 0.0~9 uDO/min is
measured. The tic complex according to the invention
thus does have a proteolytic activity wf-th respect to the
elastin.
NBA assay
(N—Bloc—L—Alaninate—para—nitrophenyl ester)
Thi s assay consists in spectrophotometrically dosing at
347.5 rm the esterase activity present in the elastase by
determiqing the itrophenol released (Vissier L. et al.
Biochim, s Acta 268 (1972) 207.260).
This dosage is not specific to an e iastolytic activity,
but is ve ry fast (kinetics of 3 min) a nd has good
sensi:ivity
ndeed, wit? 50 u" 0: complex diluted 100 times, a
varia:ion o f 0.235 uDO/min is measured. T 1e enzymatic complex
thus does nave an. activity enabling tt e release of para—
nitrophenol.
enolxtic activity assay
Azocoll assay
This assay consists in spectrophotometrically dosing at
520 nm the peptides released by the ysis of the azocoll
(ground collagen stained with a Bordeaux red dye) (Chavira,
Analytical Biochemistry 136 (1984) 446—450).
This assay sucfiers from the same drawbacks as that of
n Red (insoluble substrate), but has good
sensitivity: with 50 pl of complex diluted 50 times, a D0 of
0.398 is Heasured after incubation during 10 Hdnutes. The
enzymatic complex according to the invention thus does have a
proteolytic activity with respect :0 the azocoll.
The proteolytic activity of the enzyma:ic complex
according to the invention is thus established. The
purifiication :
0' said complex has not altered its activity.
The pro:ei1 complex thus obtained and characterized can
be ased in animal therapeutics, namely by way of an
immunostimuiant agent and as (a regeneration agent for the
Peyer’s Patches, in ular for older animals.
Additionally, the enzymatic complex enables the
regeneration of intestinal vi‘li in older animals (rats,
chickens, etc) and thus greatly improves the intestinal
tion of essential nutrimerts.
The enzymatic complex according to the present invention
is mainly intended for animal husbandry and more particularly
for the improvement of cattle feed (bovine, porcine, ovine)
and poultry, rabbit and tish teed as well as feed for any
other monogastric anima l.
Incorporated at doses from 0.025 to l g/kg to feed
compositions, this enzymatic complex enhances the metabo“ SID
0: these animals and contributes towards ensuring better
general health. Pre ferably, the doses of the complex
introduced into the feed. compositions are of 0.05 to 0.2
g/kg.
In the case of y farming, maintaining good general
health is important as this avoids the propagation of
illnesses caused by promiscuity or con:finement. The morbidity
rate is thus ntially reduced and notably the mortality
2O rate by bacterial contamination o "arr1 animals.
The feed composit ions based OH this enzyme mixture
contain one or several excipients or vehicles for nut:iments,
such as for example, grain flour (wheat, maize, rye, rice,
millet, and sorghum), n, ca rbohydrates se,
mannose), loading elements (casein, bran, cellulose,
cellulose derivatives) and/or mineral elements (chalk, clay,
bentonite, ) and all other elements used in animal
ion.
These compositions are thoroughly dry mixed with the
complex in a rOtating drum, or the enzymatic complex, in a
liquid and particular ly s form, is sprayed as a
fluidized bed on a feed composition and then granulated. Such
operations may be performed. at variable atures from
°C to 60°C.
The use 0: an tic complex that is purer and more
active thereby enables the biological ty of the
nutritive compositions according to the invention to be
better dosed and a satis:factory homogenization o: the results
to be obtained.
For this reason, feed compositions according to the
invention have an enzymatic complex t that is clearly
determined and nt.
Thus, here is a non—exhaustive list or examples 0‘ Seed
compositions intended for farm animals that contain the
enzymatic complex according to the invention mixed with
excipien:s or appropriate vehicles of nutrition.
y feed composition
A composition is ed for a batch of 10 kg adding the
enzymatic complex:
Wheat flour 2.5 kg
Lactose 4.5 kg
Bran 3 kg
Enzymatic complex according the invention 0.2 g/kg.
Such a composition is incorporated at a proportion of 0.5
kg per me':ric ton 0'‘ poultry Seed.
:y feed composition
A composition is prepared for a batch o? 20.1 kg to which
the enzymatic complex is added and which is then incorporated
into the poultcy ‘eed:
Wheat flour 1.5 kg
Oat s'our 4.5 kg
Cellulose 14 kg
Colloidal silica 0.100 kg
Enzymatic complex according the invention 0.1
g/kg.
Pig feed composition
A ition is prepared cor a batch of 10 kg to which
the enzymatic complex is added and which is then incorporated
into the pig feed:
Wheat flour 4
Bran 2
Casein 3
9otato starch 1
Enzymatic complex ing the invention 0.2
g/kg
Cattle feed composition
A composition is prepared in the same way using:
Rice starch 0.5 kg
?otato starch 2.5 kg
tic complex according the invention 0.2
g/kg.
The premix thus formed is added little by little to a
mixture 0: 6 kg of mu:ton protein powder and 14 kg of casein.
Tie homogenized preparation is intended to be incorporated
into the cat:le :eed in a proportion of 0.5 kg per metric ton
0; feed.
y _eed composition5
The composition is prepared for a batch o: 10 kg to which
the enzymatic complex is added:
Maize 5.65 kg
Soybean meal 3.23 kg
seed meal 0.3 kg
Wheat germ flour 0.4 kg
Chalk 0.13 kg
Di—calcium phosphate 0.16 kg
.aC; 0.03 kg
Amino acid complex 0.1 kg
Enzymatic complex according the invention 0.2
g/kg.
The enzymatic complex according to the ion may also
be added to o””—Lhe—shelf poultry feed compositions so as to
supplemert these preparations.
Fish feed composition
The iollowing1) composition. is prepared. and incorporated
into the Sish ‘eed to‘lowing the customary practice.
Fish meal 50.8%
Animals fats and oils 28%
Soybean meal 9.9%
Wheat flour 10%
Vitamin complex i 0\0
(A, D, E, C,m)
Enzymatic complex 0.3 %
according the invention
Experimental part
To determine the action of the complex according to the
invention on intestinal villi, assays are performed on
relatively old rats (more than seven months old).
These rats are given food based on vegetal proteins
ively of tre type wheat flour, soybean, or maize. The
enzymatic complex according’ to the invention as described
previously is added to this feed in the proportion 0“ O.’ OI
0.2 g/kg.
The results are explained in figures 1 to 4. These
figures are photonic microscopic views (enlargement x120) 0;
:he villi of the duodenum and jejunum
Figure 1 shows the duodenal villi 0: rats not having
received the enzymatic complex according to the invention in
their feed.
Figure 2 shows the al villi 0: rats having received
:eed incorporating the enzymatic complex according' to the
invention.
Figure 3 shows the jejunal vil li of rats not having
received the enzymatic complex ing to the invention in
their feed.
Figure 4 shows the jejunal villi of rats having received
feed. incorporating the tic complex according‘ to the
invention.
This establishes that the addi tion of the :ic
complex in the feed of older animals (rats, chicken, etc.)
increases the size and number of tre villi in both :he
duodenum and the jejunum. Increases C
Oi up to 70% have been
observed.
These results are erable to intestinal microvilii.
Similar results are obtained with chickens, pigs and fish.
An increase in the surface area enables faster tion
o: nutriments or medicines.
This improved absorp:ion is observed in trouts receiving
feed containing an antibiotic. In the assay, the feed
contains 4g/kg o: oxyte:racycline and the results are read
after 48 hours. An increase in the tissue concentration of
the antibiotic is thus observed of a magnitude of 100% in the
liver (5.75 instead of 2.5 microgram/g), 400% in the muscles
(1.75 d of 0.32 ram/g) and 1000% in the kidneys
(5.75 instead of 0.5 microgram/g).
Similar s are obtained with chicken and pigs.
This increase in density may lead to an increase in the
local production of physiologically active compounds such as
intestinal hormones.
Definitions of specific embodiments of the invention as
claimed herein follow.
According to a first embodiment of the invention, there
is provided use the of an enzymatic complex comprising a
mixture of proteases obtained by culturing a Streptomyces
fradiae strain to supplement farm animal feed, wherein one of
the proteases has an ctric point of around 7.0 and a
specific ty of around 150,000 Anson units/mg of protein
and another se has an isoelectric point of around 8.0
and a specific activity of around 38,000 Anson units/mg of
protein, wherein greater than 80% of the activity of said
enzymatic complex is due to these two types of proteases.
According to a second embodiment of the invention, there
is ed a feed composition for farm s comprising
the enzymatic complex according to the first embodiment and
possibly including ents or vehicles for nutriments.
According to a third ment of the invention, there
is provided a manufacturing process for the enzymatic complex
according to the first or second ment, wherein a
Streptomyces fradiae strain is cultured, in that the
fermentation broth is filtered, in that the enzymatic complex
is thereafter extracted by ultrafiltration and ion exchange
chromatography followed by an isoelectric focusing and
finally, the complex thus obtained is lyophilized.
Claims (9)
1. Use of an enzymatic complex comprising a mixture of proteases obtained by culturing a Streptomyces fradiae strain to supplement farm animal feed, n one of the proteases 5 has an isoelectric point of around 7.0 and a specific activity of around 150,000 Anson units/mg of protein and another protease has an isoelectric point of around 8.0 and a specific activity of around 38,000 Anson units/mg of protein, wherein greater than 80% of the activity of said enzymatic 10 complex is due to these two types of proteases.
2. The use according to Claim 1 as a feed supplement in powder, liquid or any other form suited to its mixture with feed compositions to improve the general condition of factory farm animals. 15
3. The use ing to Claim 2, wherein the enzymatic complex in powder form is dry mixed with a feed composition in a rotating drum until nized.
4. The use according to Claim 2, wherein the enzymatic complex in liquid form is ized in a fluidized bed onto 20 a feed composition, and is then granulated.
5. The use according to any one of Claims 1 to 4 in poultry feed.
6. The use ing to any one of Claims 1 to 4 in pig feed. 25
7. The use according to any one of Claims 1 to 4 in fish feed.
8. A feed composition for farm s comprising the enzymatic complex according to any one of Claims 1 to 4 and possibly including excipients or vehicles for nutriments. 30
9. A manufacturing process for the enzymatic complex ing to any one of Claims 1-8, wherein a Streptomyces fradiae strain is cultured, in that the fermentation broth is filtered, in that the enzymatic complex is thereafter extracted by ultrafiltration and ion exchange chromatography 35 followed by an isoelectric focusing and finally, the complex thus ed is lyophilized. ation] KEB
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR12/03171 | 2012-11-26 | ||
FR1203171A FR2998450A1 (en) | 2012-11-26 | 2012-11-26 | USE OF AN ENZYMATIC COMPLEX IN FEEDING ANIMAL BREEDING ANIMALS |
PCT/FR2013/000305 WO2014080094A1 (en) | 2012-11-26 | 2013-11-22 | Use of an enzymatic complex in the feeding of livestock animals |
Publications (2)
Publication Number | Publication Date |
---|---|
NZ708401A NZ708401A (en) | 2020-09-25 |
NZ708401B2 true NZ708401B2 (en) | 2021-01-06 |
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