What are enzymes?

2006-02093-01

Agenda
Enzymes in general
Discovery of enzymes
Production of enzymes
Enzymes for detergents

What are enzymes and what do they do
Enzymes are biological catalysts in the form
of proteins
Enzymes are present in all living cells, where they drive chemical
reactions and vital metabolic processes
Enzymes reduce resource usage and increase the efficiency of the
process
Enzymes are not consumed in the process
Enzymes operate under gentle conditions
Enzymes are fully biodegradable, and can replace chemicals in
industrial production

Enzymes as proteins
Ascorbate oxidase

Enzymes as proteins
BacteriaI aIpha amyIase
Active site residues
CaIcium ions
CaIcium ions

Enzymes as proteins

Binding Specificity
Even when different substrate molecules are present, only those that have
the specific shape complementary to the active site are able to bind with the
enzyme's active site.

Enzymes as catalysts
nzymes cataIyse reactions by Iowering the
activation energy necessary for a reaction to occur.
+ S S + P

Enzymes in general - conclusions
Chemical reactions that take place under
mild conditions
Highly specific action
Very fast reaction rates
Numerous enzymes for different tasks

Agenda
Enzymes in general Enzymes in general Enzymes in general
Discovery of enzymes
Production of enzymes Production of enzymes Production of enzymes
Enzymes for detergents Enzymes for detergents Enzymes for detergents

Enzyme discovery and screening
Found in nature

Enzyme discovery and screening
Grown in Laboratory

Screening in many ways
Growth conditions inducing
enzyme production
Growth on
selective media

Screening in many ways
Detection of enzyme
activity
High
through put

Screening process to identify relevant
enzymes
#eIevant substance
(stain) is defined by
customer
Thousands of
enzymes are appIied
to the substance to
identify
reIevant/efficient
enzyme

dentification of final enzyme

Enzyme fermentation
Fermentation to produce
industrial enzymes starts with a
vial of dried or frozen
microorganisms called a
production strain.
This production strain is selected
to produce large amounts of the
enzyme(s) of interest.

Enzyme fermentation

Agenda
Enzymes in general Enzymes in general Enzymes in general
Discovery of enzymes Discovery of enzymes Discovery of enzymes
Production of enzymes
Enzymes for detergents Enzymes for detergents Enzymes for detergents

Fermentation of enzymes
Compressor
Mixing of
nutrient medium
Water
InocuIation
fIask
Seed fermenter
Air
SteriIe fiIter Fermented broth for
recovery of enzyme
CooIing water
xhaust
SteriIisation
Raw
materials
round grain/corn
Starch
GIucose
Sugar
Soy bean meaI
GIuten
Corn steep Iiquor
Casein
Phosphates
SuIphates
Ammonium saIts
Carbohydrate
s:
Proteins:
Salts:

Recovery of enzymes
Fermenter
CuIture
broth
CooIing
UItrafiItration
StabiIisation
BacteriaI fiItration
BacteriaI
fiItration
A
B
Drum fiItration
FiItration of
enzyme
crystaIs
for granuIation
Liquid
concentrate
Liquid
product
Pretreatment
FiIter aid
is added
Preservatives
are added
CrystaIIisation
nzyme
crystaIs for
granuIation

Granulation of enzymes
#aw materiaIs
nzyme crystaIs
or Iiquid
concentrate from
the recovery pIant
GranuIation mixer
FIuid-bed dryer
Sifter
Sifter
Hopper
Fibre drums
Coating mixer
FIuid-bed cooIer
GranuIated finaI product
Processing aids
are added
Big bag

Produced in Iarge quantities

Commercial enzyme formulations
(stabilized and standardized)
Liquids
Non dusty granulates (incapsulated enzymes)
mmobilized dry products

Savinase T
Coated core, rounded beads, irreguIar shape.
Note the very smooth coating without hoIes and the wide variation in size.
0,8 mm

Novozymes granulation technology
Enzyme composition
Granulate Stability
Dust
Enzyme release
Color
Particle size distribution

PG
TiO2
KaoIin/
CaCO3
Na2SO4
CeIIuIose fibers
nzyme
Dextrin
KaoIin/CaCO3
Sugar/starch
PVA
nzyme
PVA
MgSO4
PVA
TiO2
Contrast coIour
Surfactant
T-granulate Competitor product
Comparison of granulate structure

Microscope picture
T-granulate Competitor product

SEM Pictures
T-granulate Competitor product

Set-up for mechanical stability test of granules
Microscope and camera
Weight
Glass plate
Granule

Competitor granulate
Photographs of granulate
upon increased pressure
0 g
200 g
400 g

T-granulate
Photographs of granulate
upon increased pressure
400 g
1000 g
0 g

Agenda
Enzymes in general Enzymes in general Enzymes in general
Discovery of enzymes Discovery of enzymes Discovery of enzymes
Production of enzymes Production of enzymes Production of enzymes
Enzymes for detergents

6 different enzyme classes are
used in detergents world wide
Protease
Lipase
Amylase
Cellulase

Proteases are basic ingredients
Protease
Lipase
Amylase
Cellulase
used in 75 % of all detergents
used since 1963
removes stains
basic ingredient in ADW
cannot be substituted by any other ingredients
Cocoa
Grass
Protease
no yes

Wash Performance of Savinase 6.0T
25 C, TOM, Regular powder, 10 dH, 20 min, EMPA 117 swatch
0
5
10
15
20
25
30
0 0,2 0,4 0,6 0,8 1
nzyme Concentration (%)

#
e
m
i
s
s
i
o
n

Unwashed Unwashed

Concrete examples of test/wash results
Protease effect on blood spots
not washed washed 3 times,
no protease
washed 3 times,
with protease

Concrete examples of test/wash results
No
protease
With
Protease
#e-soiIing
#e-soiIing

Protease Action
Particulate stain
Protein
Textile fiber

ParticuIate stain
Protein
TextiIe fiber
Surfactant
Protease Action

ParticuIate stain
TextiIe fiber
Surfactant
nzyme
Protease Action

ParticuIate stain
TextiIe fiber
Surfactant
Protease Action

Amylases make white whiter
available already in the sixties
often used in South America (40 % penetration)
basic ingredient in ADW
removes stains
reduces redeposition
Protease
Lipase
Amylase
Cellulase
Tomato
sauce
Chocolate
pudding
Amylase
no yes

Amylases
Amylases hydrolyse - -1,4-glucose-linkages
randomly forming soluble dextrins and
oligosaccharides
The viscosity of the gelatinized starch is
reduced and the solubility enhanced

Savinase

& Termamyl

on Cocoa Stain (EMPA 112)

38
40
42
44
46
48
50
52
54
#
e
m
i
s
s
i
o
n
40 C 60 C
Without nzyme
1 % Savinase 6.0T
1 % TermamyI 60T
0.5 % Savinase 6.0T
& 0.5 % TermamyI
60T

Removal of soils containing starch
Gelatinization and swelling are needed to make the
starch susceptible to enzymatic breakdown
For most starchy foods, various degrees of
gelatinization result from cooking
Amylases boost the removal of starch e.g. from pasta,
potato, gravy, chocolate, and baby food
Amylases prevent starch from adhering to the surface
of laundry and dishes and act as a glue for particulate
soils

Lipases give higher in-depth cleaning than
surfactants
Protease
Lipase
Amylase
Cellulase
remove fatty matter at low temperature
give in-depth cleaning
reduces redeposition
can replace surfactants
first protein engineered enzyme introduced in 1988
+ Lipase
- Lipase
Lipstick

Lipex
without Lipex
with Lipex at different dosages

Electron microscopically analysis
Longitudinal specimen of yarns soiled with olive oil
Olive oil
Unwashed
Olive oil
Washed with
detergent
Olive oil
Washed with
detergent &
lipase

Electron microscopically analysis
Cross sectional specimen of yarns soiled with olive oil
Olive oil
Unwashed
Olive oil
Washed with
detergent
Olive oil
Washed with
detergent &
lipase

Cellulases have multiple effects
available since 1985
gives multiple effects (colour care, soil removal, whitening, softening)
in top brands and special products
- ceIIuIase + ceIIuIase
MPA101
Protease
Lipase
Amylase
Cellulase

SM-picture of Fibers
Cotton, 20x wash, uropean HDP
5 g/I, 40 C, 18 dH, WITHOUT Carezyme 900 T
Cellulase effect

Cellulase effect
SEM-picture of Fibers
Cotton, 20x wash, European HDP
5 g/l, 40 C, 18 dH, WTH 1 % Carezyme 900 T

Removal of Fuzz
Clearer Colors
Slide No. 55
Novozymes Protease Products
AIcaIase: Low-medium alkalinity / wool proof
sperase: High pH & temperature tolerance
Savinase: Medium-high alkalinity
verIase: Bleach tolerance
PoIarzyme: Low-temp. & cost efficacy
Savinase UItra
AIcaIase UItra
With build-in stabiliser for Liquid detergents
Slide No. 56
Novozymes Amylase Products
TermamyI: High pH & temperature tolerance
DuramyI: Bleach tolerance
TermamyI UItra: Builder tolerance
Stainzyme: Effective on "hydrophobic and "complex starch
Slide No. 57
Novozymes Lipase Products
LipoIase: Enhanced removal of stubborn
fat stains
Lipex: Effect in 1st. wash
Slide No. 58
Novozymes Cellulase Products
CeIIuzyme: Reduces re-deposition and
greying
Carezyme: Color Care efficacy and anti-
pilling
ndoIase: Anti-redeposition effect and
improved soil removal

The End
Enzymes in general
Discovery of enzymes
Production of enzymes
Enzymes for detergents
Thank you !