Dinazyme C/S PBM

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DINAZYME
A Systems Approach to advanced enzyme technology
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Background History of enzymes
Alcoholic fermentation oldest known enzyme
reaction Y G A CO2

• 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.
• Subsequently the German chemist Eduard Buchner
  discovered (1897) that a cell-free
  extract of yeast can cause alcoholic
  fermentation.
• The ancient puzzle was then solved the yeast
  cell produces the enzyme and
  the enzyme brings about the fermentation
• 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

• Probably first experiment in which a vital
  reaction was performed outside the living
  organism
• After Buchner's discovery scientists assumed that
  fermentations and vital reactions in general were
  caused by enzymes
• Nevertheless, all attempts to isolate and
  identify their chemical nature were unsuccessful
• In 1926 the American biochemist James B. Sumner
  succeeded in isolating and crystallizing urease.

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Background History of enzymes

• Four years later pepsin and trypsin were isolated
  and crystallized by American biochemist John H.
  Northrop
• Enzymes were found to be proteins, and Northrop
  proved that the protein was actually the enzyme
  and not simply a carrier for another compound
• Research in enzyme chemistry in recent years has
  shed new light on some of the most basic
  functions of life
• Ribonuclease, a simple three-dimensional enzyme
  discovered in 1938 by American
  bacteriologist René Dubos.
• Isolated in 1946 by American chemist Moses unitz
• Synthesized by American researchers in 1969

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RIBONUCLEASE ENZYME
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Background History of enzymes

• The synthesis hooks 124 molecules in a specific
  sequence to form the macromolecule
• Led to identification of those molecular areas
  that carry out its chemical functions
• Opened up the possibility of creating specialized
  enzymes with new properties
• 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

• Non-specific reactions may result in poor product
  yields.
• High temperatures and/or pressures needed to
  drive reactions lead to high energy costs. May
  require large volumes of cooling water
  downstream.
• Harsh and hazardous processes involving high
  temperatures, pressures, acidity or alkalinity
  need high capital investment, and specially
  designed equipment and control systems.
• Unwanted by-products may prove difficult or
  costly to dispose of.
• High chemical and energy consumption, and harmful
  by-products have a negative impact on the
  environment.

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Drawbacks eliminated by enzymes

• 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?

• Six main classes by type of reaction catalyzed
• Classes are split into groups and subclasses
• Ex., lactase catalyzes the conversion of milk /
  sugar to galactose and glucose
• 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
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What are enzymes?

• 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

• If a reaction is favorable ( ?G lt 0), the
  activation energy E(act) determines how fast it
  will go.

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What are enzymes?

• 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?

• Virtually all enzymes employed in the feed
  industry are hydrolases.

• Some enzymes that are of practical value to the
  livestock industry

• Xylanases, amylases, phytases, proteases,
  cellulases, betaglucanases, and pentosanases,
  are available for use in diet formulations.

• These enzymes can be mixed and matched to form
  an enzyme cocktail to fit any particular diet
  need.

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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

• 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?

• 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.

• Most enzymes catalyze the transfer of
  electrons, atoms or functional groups.

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Factors influencing enzyme activity

• Optimum pH
• Optimum Temperature

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Factors influencing enzyme activity

• Optimum Enzyme concentration
• Optimum Substrate concentration

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Factors influencing enzyme activity

• Covalent modification

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Factors influencing enzyme activity

• Inhibitors

A competitive inhibitor
A non-competitive inhibitor molecule
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Factors influencing enzyme activity

• 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.

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Factors influencing enzyme activity

• Optimum Temperature
• Within a given range, for every 10 degrees the
  temperature increases, enzyme activity doubles.
• Enzymes become denatured at elevated
  temperatures.
• Enzymes have an optimum temperature which varies
  according to
• 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

• Normally enzymes are present in cells in low
  concentrations.
• As enzyme concentration increases the rate of the
  reaction increases linearly, because there are
  more enzyme molecules available to catalyse the
  reaction.
• At very high enzyme concentration the substrate
  concentration may become rate-limiting, so the
  rate stops increasing.

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Substrate concentration

• As the substrate concentration increases, the
  rate increases because more substrate molecules
  can collide with enzyme molecules, so more
  reactions will take place.
• 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).
• The maximum rate at infinite substrate
  concentration is called vmax,
• The substrate concentration that gives a rate of
  half the maximum rate 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

• Activity of some enzymes is controlled by others.
  
• These enzymes modify the protein chain by cutting
  it, or adding a phosphate or methyl group.
• Turns inactive enzyme into active (or vice
  versa).
• 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

• Inhibitors inhibit the activity of enzymes,
  reducing the rate of their reactions.
• Found naturally, but are also used artificially
  as drugs, pesticides and research tools.

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Inhibitors

• There are two kinds of enzymatic inhibitors.

Non-competitive
Competitive
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Competitive Inhibitors

• 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

• Inhibitor molecule is different in structure than
  the substrate molecule
• Will not fit into active site.
• Binds to another part of the enzyme molecule.
• 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.
• Reversible inhibitors - bind weakly and can be
  washed out.
• Irreversible inhibitors - bind tightly and cannot
  be washed out.
• 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
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Allosteric Effectors

• 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

• 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

• Dinazyme B/W Dry and Liquid

• Dinazyme C/S PBM Dry and Liquid

• Dinazyme PSE, (Phytase) Dry

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What is Dinazyme C/S PMB

• Supplement for corn soy based poultry and pig
  diets containing high glucan barley levels.

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What is Dinazyme C/S PMB

• 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.
  
• 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.
• 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.
• Inclusion of DINAZYME C/S in diet supplements
  provides endogenous enzymes animals lack or
  produce in low amounts. 

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WHAT MAKES DINAZYME MORE EFFECTIVE THAN OTHER
ENZYMECOMBINATIONS

• Effective action due to presence of other
  important hydrolytic enzymes, which decompose
  cellulose, lichenin, araban and pectin.
• Dinatec makes use of important technical concepts
  such as
• Covalence modification,
• The use of specific allosteric substances and
  enzyme co-factors that are conducive to higher
  enzymatic efficacy
• Enzyme concentration

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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 anti- nutritional
  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
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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

• To supplement the immature endogenous enzyme
  system.

• To maximize performance, even with limited
  digestive capacity, e.g. case of young animals
  or rapid diet changes.

• To optimize nutrient utilization of high energy
  feedstuffs.

• To enable use of normally undigestible alternate
  lower cost ingredients

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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.
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Specifications for Dinazyme C/S PBM
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GUARANTEED ANAlYSIS
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Diversified Nutri-Agri Technologies Inc.,
The End
.for now.
a Dynamic Approach to Nutri-Agri Product
Research and Technology Development