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Introduction to

detergent enzymes
8/13/2012

2009-08010-01
2 8/13/2012 NOVOZYMES PRESENTATION

CONTENT

Defining enzymes

The structure of enzymes

The various types of enzymes

Summary
3 8/13/2012 NOVOZYMES PRESENTATION

CONTENT

Defining enzymes

The structure of enzymes

The various types of enzymes

Summary
4 8/13/2012 NOVOZYMES PRESENTATION

What are enzymes?

Enzymes are:
 Proteins
 Natural catalysts
 Not living organisms
 Efficient and specific

Enzymes are natural catalysts present in all living


cells, converting nutrients into energy and
building blocks, and making new cellular
structures.

Industrial enzymes are made by microorganisms.


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Proteins are polymers of amino acids

A chain of 200-500 amino acids


Protein structure R1 R2…: 20 different side chains
polypeptide chain => 20 different common amino
acids

R1 O R3

CH NH C CH NH COOH
H2N C CH NH C CH CH

O R2 O R4 Rx

Peptide
N-terminal linkages C-terminal
amino acid
amino acid
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Structure and function

Sequence of amino acids

Structure

Function

THE FUNCTION
DEPENDS UPON
THE STRUCTURE
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Protein structure overview


 Primary structure
 Sequence of amino acids in the protein chain
 Secondary structure
 Interaction between neighboring amino acids to form helical
structure (alpha or beta)
 Tertiary structure
 Folding of helical and non-helical(coiled) parts of the protein to
form a compact structure also called globular protein
 Most enzymes are globular proteins
 Quaternary structure
 Aggregates of individual protein chains
 These individual proteins can be identical or they can have
different structures (example: hemoglobin)

THERE ARE FOUR STRUCTURAL


ENZYMES ARE GLOBULAR PROTEINS LEVELS OF PROTEINS
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CONTENT

Defining enzymes

The structure of enzymes

The various types of enzymes

Summary
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Denaturing of proteins

AMINO ACIDS
• PROTEINS
ARE LOCATED NEAR DENATURE IN
ONE ANOTHER IN
THE ACTIVE FORM
EXTREME
BUT NOT IN THE CONDITIONS LIKE
DENATURED FORM.
CO FACTORS HELP
HIGH TEMPERATURE
STABILIZE (FRYING AN EGG)
STRUCTURE - METAL
IONS, CO-ENZYMES,
AND IN THE
(CALCIUM) PRESENCE OF
CERTAIN HARSH
CHEMICALS (WITH
HIGH PH AND
BLEACH)
Substrate
S
• WHEN AN ENZYME
DENATURES, LOSES
ITS STRUCTURE, IT
ALSO LOSES ITS
FUNCTION
10 8/13/2012 NOVOZYMES PRESENTATION

Enzyme inactivation

Inhibition of enzyme activity


 Refers to partial or complete loss of
enzymatic activity by a non-
substrate compound (inhibitor)
 Inhibition is usually due to binding
of the inhibitor by the enzyme, Inactivation…denaturation
which can occur at the active site  Refers to irreversible loss of catalytic
or another place on the enzyme activity of an enzyme
 Inhibition can be reversible or  Occurs when the active site is
irreversible distorted or destroyed due to partial
 Irreversible inhibition leads to or complete unfolding of the protein
complete loss of activity  Usually caused by extreme pH, high
(inactivation) temperature, high salt levels, and
specific denaturing agents
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Benefits of detergent enzymes

 Boost the performance of a basic detergent


 Completely remove spots and stains are
removed
 Maintain fabric whiteness
 Maintain colored fabrics
 Improve removal of visible contaminations
and of non-visible contaminations that
increase soil binding onto the fabrics and
cause visible staining
 Reduce formulation costs and increase
wash performance by replacing surfactants
and builders with enzymes
 Reduce the environmental impact of
washing
Electron microscopic analysis

Olive oil unwashed Olive oil washed with Olive oil washed with
detergent detergent and lipase
Oil removal from the inside of cotton fibres

Olive oil unwashed Olive oil washed with Olive oil washed with
detergent detergent and lipase
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Enzymatic catalysis

Substrate ENZYMATIC
Enzyme-substrate complex REACTION
transition state

Enzyme

Products

Enzyme + products
Enzyme

THE ENZYME BREAKSDOWN THE SUBSTRATE


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Enzyme classification
Classification Reaction catalyzed

Oxidoreductases Oxidation-reduction (e-transfer)

Transferases Transfer of functional groups

Hydrolases Hydrolysis (transfer of functional groups to


water)
Lyases Addition of groups to double bonds, or the
reverse
Isomerases Isomerization (transfer of groups within
molecule to yield isomeric forms)
Ligases Formation of C-C, C-S, C-O & C-N bonds by
condensation reactions coupled to ATP
hydrolysis/cleavage
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The most common detergent enzyme types


Enzyme type Substrate Stain example

Protease Proteins Blood, egg, grass, dairy


products
Amylase Starch Cocoa, pasta, fruit, potatoes,
oatmeal, gravy, tomato
sauce, chocolate
Lipase Triglycerides Fats, vegetable oils

Cellulase Cellulose Cotton

Mannanase Mannans/gums Chocolate sauce

Organic compounds are hydrolyzed by enzymes in detergents


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Commercially available detergent enzymes


from Novozymes
Enzyme type Commercial name Benefits
Protease Savinase, Everlase**, Protein stain removal
Esperase, Alcalase,
Polarzyme, Ultra
products
Amylase Termamyl, Duramyl, Starch stain removal
Stainzyme, Stainzyme
Plus**, Termamyl
Ultra
Lipase Lipolase, Lipex Grease removal
Cellulase Celluclean, Celluclean Whiteness and color
Classic, Carezyme, care
Carezyme Premium
Mannanase Mannaway Mannan stain removal
Available in granulate and liquid forms
Typical dose: 0.4-2% in HDP; 0.2 to 1.0% in HDL
** only the granule version is bleach stable in the formulation
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Development of detergent enzyme granulates


Year Physical form Appearance Diameter, mean µm

<1968-69 Powder ~ 30

1970 Prill ~ 450

1972 Marumes ~ 600-700

1981 T granulate ~ 500-600

1995 T granulate type W ~ 500-600


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CONTENT

Defining enzymes

The structure of enzymes

The various types of enzymes

Summary
20 8/13/2012 NOVOZYMES PRESENTATION

Proteases

HYDROLYZES PROTEIN AMIDE


BONDS

REMOVES THE STIFF AND DULL


GUM LAYER FROM THE RAW
SILK FIBER FOR IMPROVED
LUSTER AND SOFTNESS

MODIFIES THE SURFACE OF


WOOL AND SILK FIBERS OR
FABRICS FOR NEW AND UNIQUE
HANDS

REMOVES BLOOD, EGG, GRASS,


AND DAIRY PRODUCTS STAINS
21 LUNA 2006-42711-01

Enzymatic breakdown of proteins

R1 O Rn-1 O Rn O

H3N CH C NH CH C NH CH C O

H2O
endoprotease exoprotease

R1 O Rn-1 O Rn O

H3N CH C NH CH C O H3N CH C O
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Better cleaning with Polarzyme®

POLARZYME
OFFERS
SIGNIFICANT
OVERALL
CLEANING
BENEFITS BY
BREAKING
DOWN
Without Polarzyme With Polarzyme PROTEIN
STAINS AND
ELIMINATING
PROTEIN
REMAINS ON
GARMENTS
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Wash performance of Savinase® 6.0T on blood

Unwashed Improvement in wash performance with increasing enzyme concentration

25°C, TOM, REGULAR POWDER, 10°DH, 20 MIN, EMPA 117 SWATCH


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Amylases

HYDROLYZES -1,4-LINKAGES IN
STARCH

IS SPECIFIC TOWARDS STARCH


AND REMOVES THE STARCH
WITHOUT CAUSING ANY FIBER
OR FABRIC DAMAGE

EXAMPLES OF STARCH BASED


SOILS: COCOA, PASTA, FRUIT,
POTATOES, OATMEAL, GRAVIES,
TOMATO SAUCES, CHOCOLATE
Enzymatic breakdown of starch

AMYLASE
CONVERTS
INSOLUBLE
STARCH TO
Amylopectin SMALL
WATER-
Amylose SOLUBLE
FRAGMENTS

Amylase
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Stainzyme® can remove a lot of common


food stains

EXAMPLES OF
STAINS
REMOVED BY
STAINZYME
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Stainzyme® offers better stain removal


Without Stainzyme With Stainzyme

AMYLASES LIKE STAINZYME CAN REMOVE


A LOT OF COMMON STARCH AND FOOD STAINS
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Enzymes eliminate dinginess caused by starch residues

Test item:
Cotton sports socks,
With amylase
spotted with 50mg starch

Washing:
4 wash cycles, machine wash

Soiling:
With tennis court clay,
between wash cycles

Without amylase
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Lipases

LIPASES ASSIST IN
THE REMOVAL OF
TRIGLYCERIDE-BASED
SOILS

EXAMPLES OF SUCH
STAINS ARE TALLOW,
SIZE LUBRICANTS,
FATS, AND
VEGETABLE OILS
30 LUNA 2006-42711-01

Lipase converts insoluble fats to water-


soluble fragments

Hydroxyl group

Fatty acyl group

Triglyceride Diglycerides Monoglycerides Glycerol

LIPASE HYDROLYSIS OF TRIGLYCERIDES AND ASSISTS


IN THE REMOVAL OF TRIGLYCERIDE-BASED SOILS
31 8/13/2012 NOVOZYMES PRESENTATION

Lipolysis during wash

LIPOLYSIS
DURING WASH
LIPEX® IN PH-
STAT ASSAY
W. COTTON
AND COTTON
ELASTIN
SWATCHES
WITH 200 L
LARD SOIL,
Lipex

1.8 G/L US
HDL, 150 PPM
CA/MG 2:1
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Combined wash and anti-redeposition effects of lipase


Unwashed No lipase
textiles

With lipase

LARD/CARBON BLACK ON COTTON, LIPSTICK SWATCHES, AND CLEAN POLYESTER


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Lipase sensitive natural stains

Without lipase With lipase


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Cellulases

HYDROLYZES THE B-1,4-


LINKAGES IN CELLULOSE

MODIFIES THE SURFACE OF


CELLULOSIC FIBERS AND FABRICS
TO ACHIEVE A DESIRED SURFACE
EFFECT

EXAMPLES
• BIO-POLISHING OF COTTON
KNIT TO PREVENT PILLING
• COLOR BRIGHTENING (COLOR
CLARIFICATION)
• SOIL RELEASE (SOIL
REMOVAL)
• SOFTENING
• WHITENING (ANTI SOIL
REDEPOSITION)
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Structure of cellulases

Cellulase

CELLULASES HAVE THE PRIME EFFECT ON THE FABRIC - AND THAT IS TRUE FOR ALL
FABRICS CONTAINING CELLULOSE , FOR EXAMPLE COTTON
THE EFFECT OF CELLULASE ON A MOLECULAR LEVEL IS THE CLEAVAGE OF THE
BETA-1,4 BOND OF CELLULOSE
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Benefits of cellulase

• COLOR
BRIGHTENING /
CLARIFICATION
With cellulase Without cellulase
• SOIL RELEASE
• SOFTENING
• WHITENESS DUE
TO ANTI-
REDEPOSITION
37 8/13/2012 NOVOZYMES PRESENTATION

Anti-pilling and anti-redeposition effects of


cellulase

without cellulase with cellulase

ANTI-PILLING AND ANTI-


REDEPOSITION BENEFITS ON
WHITE COTTON FABRIC AFTER
MULTIPLE WASH CYCLES:

• LESS PILLING, FABRIC


LOOKS NEW

• LESS ANTI-REDEPOSITION,
FABRIC LOOKS CLEAN

Without cellulase With cellulase


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Get better whiteness with enzymes

Washed without enzyme Washed with enzyme


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Mannanase

MANNANASES ARE
CARBOHYDRASES REMOVING
MANNAN – ALSO CALLED GUMS

MANNAN IS USED WORLDWIDE


AS THICKENER AND STABILIZER
IN SAUCES, SALAD DRESSINGS,
ICE CREAMS AND DESSERTS,
CHEESES, INSTANT NOODLES,
PET FOODS, BREAD, AND
BEVERAGES – AND ALSO IN
TOOTHPASTES, SHAMPOOS,
AND CONDITIONERS
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What is mannan?

Mannans are
polysaccharides made of
mannose and galactose

Mannose residues are


linked together by β-1,4-
linkages with side chains
of galactose attached to
the backbone by α-1,6-
linkages

Mannanases cleaves the β-


1,4-linkages

TWO MAJOR TYPES OF GALACTOMANNANS:


GUAR GUM (E412)
LOCUST BEAN GUM (E410)
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Effect of mannanases

MANNANASE
REMOVES
THE STAIN

Mannanase

Mannan on cotton Clean cotton

.
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Mannaway® visibly removes mannan stains


Unwashed Washed wo. Washed w.
Mannaway Mannaway

Chocolate
milkshake
(Equest)

ASDA chocolate ice


cream
(Equest)

Sainsbury´s
chocolate ice cream
(Equest)

Salad dressing
(CFT CS-6)
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CONTENT

Defining enzymes

The structure of enzymes

The various types of enzymes

Summary
44 8/13/2012 NOVOZYMES PRESENTATION

In summary

• Enzymes are biological


catalysts in the form of
proteins and not living
organisms
• Their specific function
depends on their structure
• Most detergent enzymes are
hydrolases (protease,
amylase, lipase, cellulase)
• Each enzyme has a specific
range of operating
parameters for its optimum
performance
• Industrial enzymes are
made by micro-organisms
• Enzymes are readily
biodegradable .
45 8/13/2012 NOVOZYMES PRESENTATION

THANK YOU

YOU CAN LEARN MORE ABOUT


NOVOZYMES’ DETERGENT ENZYMES AT
WWW.NOVOZYMES.COM/DETERGENT