Download Dinazyme C/S

DINATEC
A Systems Approach to advanced enzyme technology
DINATEC
Background History of enzymes
Alcoholic fermentation oldest known enzyme reaction
Y+G
A + CO2
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Phenomena believed to be spontaneous reactions until 1857,
when the French chemist Louis Pasteur proved that fermentation
occurs only in the presence of living cells.
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Subsequently the German chemist Eduard Buchner discovered
(1897) that a cell-free extract of yeast can cause alcoholic
fermentation.
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The ancient puzzle was then solved; the yeast cell produces
the enzyme and the enzyme brings about the fermentation
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As early as 1783 the Italian biologist Lazzaro Spallanzani had
observed that meat could be digested by gastric juices extracted
from hawks.
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Background History of enzymes
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Probably first experiment in which a vital reaction was
performed outside the living organism
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After Buchner's discovery scientists assumed that fermentations
and vital reactions in general were caused by enzymes
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Nevertheless, all attempts to isolate and identify their chemical
nature were unsuccessful
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In 1926 the American biochemist James B. Sumner
succeeded in isolating and crystallizing urease.
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Background History of enzymes
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Four years later pepsin and trypsin were isolated and
crystallized by American biochemist John H. Northrop
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Enzymes were found to be proteins, and Northrop proved that
the protein was actually the enzyme and not simply a carrier
for another compound
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Research in enzyme chemistry in recent years has shed new
light on some of the most basic functions of life
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Ribonuclease, a simple three-dimensional enzyme
discovered in 1938 by American bacteriologist René
Dubos.
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Isolated in 1946 by American chemist Moses unitz
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Synthesized by American researchers in 1969
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RIBONUCLEASE ENZYME
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Background History of enzymes
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The synthesis hooks 124 molecules in a specific
sequence to form the macromolecule
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Led to identification of those molecular areas that
carry out its chemical functions
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Opened up the possibility of creating specialized
enzymes with new properties
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This potential has been greatly expanded in recent
years by genetic engineering techniques that have
made it possible to produce some enzymes in great
quantity
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How are enzymes manufactured?
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How are enzymes manufactured?
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Industry drawbacks
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Non-specific reactions may result in poor product yields.
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High temperatures and/or pressures needed to drive reactions lead to
high energy costs. May require large volumes of cooling water
downstream.
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Harsh and hazardous processes involving high temperatures, pressures,
acidity or alkalinity need high capital investment, and specially designed
equipment and control systems.
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Unwanted by-products may prove difficult or costly to dispose of.
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High chemical and energy consumption, and harmful by-products have a
negative impact on the environment.
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Drawbacks eliminated by enzymes
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Reactions carried out under mild conditions
Highly specific
Involve very fast reaction rates
Reactions are carried out by numerous enzymes with different roles.
Industrial enzymes originate from biological systems which contribute to
sustainable development through being isolated from microorganisms which
are fermented using primarily renewable resources.
Small amounts of enzymes are required to carry out chemical reactions
Reaquires little storage space.
uncomplicated and widely available equipment can be used
Reactions are easily controlled and can be stopped when the desired degree
of substrate conversion has been achieved.
Reduce the impact of collateral damage on the environment by reducing the
consumption of chemicals and energy, and the subsequent generation of
waste.
Developments in genetic and protein engineering have led to improvements
in the stability, economy, specificity and overall application potential of
industrial enzymes.
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What are enzymes?
An enzyme is a protein which
acts as a specific biological
catalyst facilitating a given
reaction by lowering the
amount of required energy.
To date, scientists have
identified over 1,500
different enzymes.
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What are enzymes?
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Six main classes by type of reaction catalyzed
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Classes are split into groups and subclasses
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Ex., lactase catalyzes the conversion of milk / sugar to
galactose and glucose
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Lactase has the systematic name beta-D-galactoside
galactohydrolase, and the classification number EC 3.2.1.23.
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SIX MAIN ENZYME CLASSES
CLASS
REACTION PROFILE
1: Oxidoreductases
Involves movement of electrons from one molecule to another. In biological systems we
usually see the removal of hydrogen from the substrate. Enzymes in this class are called
dehydrogenases. Ex., alcohol dehydrogen-ase catalyzes reactions of the type R-CH2OH + A
→ R-CHO + H2A, where A is an acceptor molecule. If A is oxygen, the relevant enzymes are
called oxidases; if A is hydrogen peroxide, the relevant enzymes are called peroxidases.
2: Transferases
This class of enzymes catalyzes the transfer of groups of atoms (radicals) from one molecule to another. Aminotransferases or transaminases promote the transfer of an amino group
from one amino acid to an alpha-keto-acid.
3: Hydrolases
Hydrolases catalyze reactions between a substrate and water, and bind water to certain
molecules. In this way larger molecules are broken up into smaller units. This class of
enzymes catalyzes the cleavage of peptide bonds in proteins, glucosidic bonds in
carbohydrates, and ester bonds in lipids.
4: Lyases
Lyases catalyze the addition of groups to double bonds or the formation of double bonds
through the removal of groups. Thus bonds are cleaved using a different principle to
hydrolysis. Pectate lyases, for example, split the glycosidic linkages by beta-elimination.
5: Isomerases
Isomerases catalyze the transfer of groups from one position to another on the same
molecule. These enzymes change the structure of a substrate by rearranging its atoms.
6: Ligases
Ligases join molecules together with covalent bonds. These enzymes participate in
biosynthetic reactions where new groups of bonds are formed. Such reactions require the
input of energy in the form of co-factors such as ATP.
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What are enzymes?
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Globular, water soluble proteins, (few exceptions)
Allows / facilitates chemical reactions to occur such as
those that release nutrients from feed during digestion
Without the enzyme catalyst the reaction would
either not take place or would happen very slowly
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If a reaction is favorable ( ∆G < 0), the activation
energy E(act) determines how fast it will go.
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What are enzymes?
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Though an enzymatic catalyst takes part in the
chemical reaction it remains unchanged and is
available to repeat the task
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What are the most important enzymes to our industry?
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Virtually all enzymes employed in the feed industry are
hydrolases.
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Some enzymes that are of practical value to the
livestock industry:
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Xylanases, amylases, phytases, proteases, cellulases,
betaglucanases, and pentosanases, are available for
use in diet formulations.
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These enzymes can be mixed and matched to form
an enzyme cocktail to fit any particular diet need.
DINATEC
Why are enzymes needed in feed formulations?
Trials confirm that enzyme supplementation results in improved
animal performance.
 Young animals lack many endogenous enzymes or sufficient
quantities thereoff.
 Sick animals may have a damaged intestinal lumen resulting in
limited nutrient absorption.
 Animals under stress or at a high level of production may have
an impaired digestive system.
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Why are enzymes needed in feed formulations?
Problems in feed ingredients:
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Raw materials may contain anti-nutritive factors. Ex. pentosans
or betaglucans present in wheat or barley.
 Addition of appropriate enzyme aids digestion of the material
improving feed value.
 Increasing environmental awareness and restrictions on
pollutants and contaminants confirm the value of enzymes in
the breakdown of such materials. Ex Phytase/ phosphorus
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How do enzymes work?
Specificity
 Specific enzymes may be incorporated into specific diets in
order to solve specific problems
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How do enzymes work?
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Enzyme catalyzed reactions are often
from 100 million to more than 10 billion
times faster than the same reaction in
the absence of the enzyme.
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Most enzymes catalyze the transfer of
electrons, atoms or functional groups.
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Factors influencing enzyme activity
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Optimum pH
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Optimum Temperature
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Factors influencing enzyme activity
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Optimum Enzyme concentration
Optimum Substrate concentration
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Factors influencing enzyme activity
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Covalent modification
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Factors influencing enzyme activity
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Inhibitors
A competitive inhibitor
A non-competitive inhibitor molecule
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Factors influencing enzyme activity
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Allosteric Effectors
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Factors influencing enzyme activity
 Optimum
pH: pH at which enzymes
operate best. Activity decreases on either side
of pH optimum.
DINATEC
Factors influencing enzyme activity
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Optimum Temperature:
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Within a given range, for every 10 degrees the temperature increases, enzyme activity doubles.
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Enzymes become denatured at elevated temperatures.
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Enzymes have an optimum temperature which varies according to:
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Enzyme source.
Salt levels in the medium to which the enzyme is added. (For example, amylases from
animal sources are less heat stable than those from fungal sources (Aspergillus) which are
in turn less stable than bacterial amylases (Bacillus).
Mineral Content: Certain minerals stabilize enzymes while others cause inactivation.
Calcium and magnesium are essential for good starch breakdown (amylases) and increase
enzyme stability to temperature. Heavy metals such as iron are typically detrimental to
enzymes, and may in some cases be used to inactivate or stop enzyme reactions.
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Enzyme concentration
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Normally enzymes are present in cells in low
concentrations.
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As enzyme concentration increases the rate of the
reaction increases linearly, because there are more
enzyme molecules available to catalyse the reaction.
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At very high enzyme concentration the substrate
concentration may become rate-limiting, so the rate
stops increasing.
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Substrate concentration
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As the substrate concentration increases, the rate increases because more
substrate molecules can collide with enzyme molecules, so more reactions
will take place.
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As substrate concentration gets higher the enzyme molecules become
saturated so there are few free enzyme molecules. Adding more substrate
doesn't make much difference (though it will increase the rate of E-S
collisions).
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The maximum rate at infinite substrate concentration is called
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The substrate concentration that gives a rate of half the maximum rate
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vmax,
vmax is called KM.
The vmax and KM values are useful for characterising an enzyme.
A good enzyme has a high vmax and a low KM.
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Substrate concentration
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Covalent modification
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Activity of some enzymes is controlled by others.
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These enzymes modify the protein chain by cutting it, or adding
a phosphate or methyl group.
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Turns inactive enzyme into active (or vice versa).
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Used to control many metabolic enzymes and to switch on
enzymes in the gut e.g. hydrochloric acid in stomach activates
pepsin activates rennin.
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Inhibitors
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Inhibitors inhibit the activity of enzymes,
reducing the rate of their reactions.
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Found naturally, but are also used artificially as
drugs, pesticides and research tools.
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Inhibitors
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There are two kinds of enzymatic inhibitors.
Competitive
Non-competitive
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Competitive Inhibitors
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Molecule has similar structure to normal substrate
molecule. Fits into active site of the enzyme.
Competes with substrate for the active site, so
reaction is slower.
Increase KM for enzyme, but no effect on vmax.
The rate can approach a normality if substrate
concentration is increased sufficiently.
The sulphonamide anti-bacterial drugs are
examples of competitive inhibitors.
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Non-competitive inhibitors
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Inhibitor molecule is different in structure than the substrate molecule
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Will not fit into active site.
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Binds to another part of the enzyme molecule.
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Change enzyme and active site shape so it no longer binds substrate
molecules. Result is reduction of active enzyme numbers (just like
decreasing the enzyme concentration). Therefore decrease vmax, but
have no effect on KM.
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Reversible inhibitors - bind weakly and can be washed out.
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Irreversible inhibitors - bind tightly and cannot be washed out.
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Poisons like cyanide, heavy metal ions and some insecticides are all
examples of non-competitive inhibitors.
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RATE EQUATION FOR PRODUCT INHIBITION
Michaelis-Menten equation
RATE EQUATION
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Allosteric Effectors
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Activity of some enzymes is
controlled by certain molecules
binding to a specific regulatory or
allosteric site on the enzyme.
Allosteric site is distinct from the active site
Different molecules can inhibit or activate the enzyme,
allowing sophisticated control of the reaction rate
Few enzymes can do this. They are often at the start of long
biochemical pathways
Generally activated by the substrate of the pathway and
inhibited by the product of the pathway, thus only turning the
pathway on when it is needed
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Allosteric Effectors
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Economic benefits
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Increases daily weight gain
Increases egg production
Lowers feed conversion
More uniform weights / increased nutrient absorption
Lower incidence of digestive problems caused by
unassimilated fiber which also improves litter quality
Reduces fecal volume and nitrogen excretion levels
Cleaner eggs and better egg yolk color
Use of lower cost ingredients
Maintains and improves performance levels
Increases ratio of lean to fat tissue
Can "inactivate" mycotoxins in feeds
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What is Dinazyme?
Several types of enzyme technologies are offered as Dinazyme
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Dinazyme B/W Dry and Liquid
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Dinazyme C/S PBM Dry and Liquid
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Dinazyme PSE, (Phytase) Dry
DINATEC
What is Dinazyme C/S PMB
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Supplement for corn soy based
poultry and pig diets containing
high glucan barley levels.
DINATEC
What is Dinazyme C/S PMB
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A diet supplement which enhances nitrogen utilization and increases
protein digestibility with the active ingredient protease, resulting in
increased absorption of amino acids and peptides.
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DINAZYME C/S® also contains amylase-breaks down starch content
and Xylanase, a complex hydrolytic enzyme preparation which has an
effect on hemicellulose substrates containing xylan, manan and glucan.
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A combination of amylase, Xylanase and protease boosts the
digestibility of typical corn and soybean meal-based diets, resulting in
more nutrients available for growth.
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Inclusion of DINAZYME C/S® in diet supplements provides endogenous
enzymes animals lack or produce in low amounts.
DINATEC
WHAT MAKES DINAZYME MORE EFFECTIVE THAN OTHER
ENZYMECOMBINATIONS
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Effective action due to presence of other important
hydrolytic enzymes, which decompose cellulose,
lichenin, araban and pectin.
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Dinatec makes use of important technical concepts
such as:
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Covalence modification,
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The use of specific allosteric substances and enzyme
co-factors that are conducive to higher enzymatic
efficacy
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Enzyme concentration
DINATEC
What can Dinazyme C/S PMB do for you?
Contents / effects:
• Protease, enhanced nitrogen utilization and increased protein digestibility
• Amylase, increased digestibility of starch in pig and poultry diets
• Betaglucanase, reduced digesta viscosity in poultry diets; decreases antinutritional effects of NSP*; reduces soluble NSP in disgesta.
Pectinase, more complete hydrolysis (digestion) of pectins in wheat and corn
based diets
Xylanase reduces digesta viscosity; decreases anti-nutritional effects of
NSP; reduces soluble NSP in digesta hence increased absorption of amino
acids and peptides.
* Non Starch Polysaccharide
DINATEC
Why should you use Dinazyme?
High energy diets high on starch and protein content are
desirable at an early age for the monogastric.
Young animal's endogenous enzyme system not fully developed.
Unable to adapt quickly enough for demands of current feed
management programs.
Immature pancreas needs time to adapt to new diet and produce
necessary amounts and types of digestive enzymes.
Result - Undigested feed is wasted.
Undigested feed promotes "nutritional scours" and provides
substrate for the growth of diarrhea-causing pathogens.
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Why should you use Dinazyme?
The Solution… Dinazyme C/S-PMB
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To supplement the immature endogenous enzyme system.
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To maximize performance, even with limited digestive capacity,
e.g. case of young animals or rapid diet changes.
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To optimize nutrient utilization of high energy feedstuffs.
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To enable use of normally undigestible alternate lower cost
ingredients
DINATEC
ECONOMICS
Data suggest that an average improvement in nutrient utilization
of 3 – 5% can be obtained
Leeson and Summers (1976) reported that high moisture
content corn harvest necessitated high temperature drying &
time retention conditions, reduced ME value of corn by as much
as 3% compared to the expected value.
DINATEC
Specifications for Dinazyme C/S PBM
EFFICACY
Effective over a wide pH and
temperature range.
PACKAGING
Available in 5, 10 and 20 kg pails or 55
lb. bags.
STORAGE
In dry location. Do not exceed 26°C.
SHELF LIFE
One year in original sealed container.
Three months in stored premixes and
feedstuffs.
USE & DOSE
For any corn/soy based feed.
Layer/Breeder feeds. 200 grms/metric
ton, Broiler feeds. 250 grms/metric ton.
Pig diets. 300 grms/metric ton.
DINATEC
GUARANTEED ANAlYSIS
ENZYME TYPE
Amylase
ECC IUB # 3.2.1.1
CONTENT U/GRAM
1600
Protease
ECC IUB # 3.4.24.28
16000
Xylanase
ECC IUB # 3. 2.1.8
1200
Pectinase
ECC IUB # 3. 2.1.8
120
Betaglucanase
ECC IUB # 3. 2.1.8
600
DINATEC
Diversified Nutri-Agri Technologies Inc.,
The End
…….…for now…….
“a Dynamic Approach to Nutri-Agri Product Research and Technology Development”