1. Tips on how best to use enzymes in your formulations
4/15/2017

2. Test methodology and examples
4/15/2017
NOVOZYMES PRESENTATION
content
Formulation advice
Test methodology and examples
Industrial applications
Summary

3. Composition examples of household powder detergents
4/15/2017
NOVOZYMES PRESENTATION
Composition examples of household powder detergents
Ingredient
Laundry Detergent
Dishwashing Detergent
Anionic Surfactants
5 - 18% --
Nonionic Surfactants
2 - 8%
1-5%
Soap
1 - 5%
Builders (zeolite, carbonates, silicates) %
32-90%
Bleaches (perborate/activator)
0 - 30%
3-15%
Polymers
0 - 5%
5-10%
Enzymes: protease
amylase
lipase
cellulase
Mannaway % % % %
0.2 – 0.5%
1-1-5%
1-2%
Fillers (sodium sulfate)
0 - 20%
Water + others
to 100%

4. Composition examples of household liquid detergents
4/15/2017
NOVOZYMES PRESENTATION
Composition examples of household liquid detergents
Ingredient
Laundry Detergent
Prespotter
Anionic Surfactants
5 - 20%
0 - 5%
Nonionic Surfactants
3 - 15%
4 - 12%
Soap
0 - 12% --
Coupling Agents (xylene sulfonate)
Builders (citrate, carbonate)
2 - 8%
0 - 2%
Buffers (triethanolamine)
Enzymes: protease
amylase
lipase % % %
Enzyme Stabilizers
0 - 8%
Water & other
to 100%

5. Formulation guidelines
4/15/2017
NOVOZYMES PRESENTATION
Formulation guidelines .
Anionic surfactant system (charged surfactants)
Negative influence on enzyme stability
AES = None > MES > AOS ³ LAS > Soap
Non-ionic surfactant system
AEO6 > NP12 > NP4 > None
Amides and Alkyl polyglucosides are ok
Recommendation
Preferably use non-ionic surfactants
Or use a combination of nonionic and anionic surfactants

6. 4/15/2017
NOVOZYMES PRESENTATION
Compatibility of enzymes with materials found in typical detergent formulations
Materials
Proteases
Amylases
Lipases
Cellulases
Enzymes
May self-hydrolyse; other
have no effect
Need stabilizer when proteases present; otherwise ok in general
Anionic surfactants
Best: AES, MES. Other: AOS and LAS tend to interact unfavorably with enzymes.
Nonionic surfactants
Ok in general.
High level may reduce
effect. Best if
anionic/nonionic ratio of
¼- ¾
Best if anionic/nonionic
ratio of ¼ - ¾
Cationic surfactants
May decrease stability
Coupling agents /
hydrotrope
MPG best. Others ok in
General, SXS level low
general
MPG best; others ok in
Buffers (prefer pH <
9 in liquids)
Ok to use Borate, TEA or
MEA. Use <5%w TEA
Keep pH at for optimal stability.
Recommended Stabilizers
Borate, MPG, sodium
formate (low pH). Calcium
may help
Calcium (e.g. CaCl2) best.
Borate, MPG, or sodium
formate when protease
present.
present
Builders
Ok in general except for strong chelating agents (e.g.
EDTA, TKPP, NTA). Keep pH at for optimal
stability
Ok in general. Keep pH at for optimal stability
Preservatives
Most ok (e.g. Proxel, Kathon, Dowicide, etc.) except formaldehyde
Bleach
Percarbonate or perborate ok to use in powders. Not recommended overall in liquids. Chlorine containing type not
recommended at all
Other Comments
Keep water content between % for optimal stability in liquids; keep powders from adsorbing moisture for
optimal stability (may use moisture barriers)
Recommendations are based on shelf-stability of enzymes in liquid formulations unless otherwise stated

7. Factors influencing enzyme stability in a liquid detergent
4/15/2017
NOVOZYMES PRESENTATION
Factors influencing enzyme stability in a liquid detergent
Water content pH
Type and amount of enzyme stabilizers
Time
Temperature
Enzyme dose
Major components such as surfactants, builders
Other components in the reaction mix (e.g., preservative, bleach)
These factors interact and each enzyme has a
specific range of operating parameters for its
optimum performance.

8. What does shelf stability mean?
4/15/2017
NOVOZYMES PRESENTATION
What does shelf stability mean?
Stability of individual enzymes in a detergent, especially liquid
Stability of enzymes in the presence of protease

9. Enzyme stability depends upon conditions
4/15/2017
NOVOZYMES PRESENTATION
Enzyme stability depends upon conditions .
1 year
1 hour
16 weeks

10. Guidelines to improve enzyme stability in formulations
4/15/2017
NOVOZYMES PRESENTATION
Guidelines to improve enzyme stability in formulations
Use stabilizers
CaCl2.2H2O to ~0.08 % w – protease and amylase stability
borate + mono propylene glycol – protease inhibitors
sodium formate – protease inhibitor especially at pH < 8
4-FPBA (4-formyl phenyl boronic acid) – protease inhibitor
Moderate the water content to 50-60% w
Lower the anionic surfactant content, especially LAS (AS)
Examine antagonistic detergent components (e.g. bleach, chelating agents)
Adjust pH to 9 or less , best if 7.5 to 8.5
Use the Novozymes’ Ultra proteases containing 4-FPBA
Alcohols/hydroxy groups stabilize proteases & facilitate solubilization of surfactants in water phase
Propylene-glycol > Ethanol >> Sorbitol > None
Acetamide, glycine, maltodextrin may be added to improve enzymatic stability.
Dosage: % (w/w)
Not compatible: Glutamic acid (Phase sep.)

11. 4-FPBA - highly effective protease inhibitor reference
4/15/2017
NOVOZYMES PRESENTATION
4-FPBA - highly effective protease inhibitor reference
reference: Inform 13 p.731 (2002)
Molecular structure of 4-Formylphenyl Boronic Acid
(4FPBA)
C=O
H-O B H
Highly efficient protease inhibitor compared to borax/propylene glycol combination
Since 4FPBA is already added to the liquid protease, added stabilizers are not needed or are greatly reduced, with minor formulation adjustments including lowering of pH
Cost-savings on added stabilizers
Good stability of protease and other enzymes in the presence of a protease
Savinase® Ultra and Alcalase® Ultra are now available

12. 4-FPBA effect in typical US model detergent (pH 7)
4/15/2017
NOVOZYMES PRESENTATION
4-FPBA effect in typical US model detergent (pH 7) 25 50 75
100 7 14 21
Days
Residual Activity (%)
No protease
Savinase 16L
Savinase Ultra 16L 25 50 75
100 7 14 21
Days
Residual Activity (%)
Savinase 16L
Savinase Ultra 16L .
incubation at 30°C

13. Stability of Savinase® at different pH values
4/15/2017
NOVOZYMES PRESENTATION
Stability of Savinase® at different pH values
The pH of the liquid detergent can also affect the stabilization of enzyme activities as illustrated in Slide 15. At increasing pH values from 9 to 10, there is a drastic decrease in residual activity for Savinase including 4-FPBA from 65% to 8%. With no inhibitor present, the decrease is from 15% to 8% revealing effect of 4-FPBA at pH but not at pH 10. Other stabilizers in addition to 4-FPBA will be needed to achieve acceptable stability at the higher pH values. Experience has shown that the level of additional stabilizer such as boric acid is not as high as used if no 4-FPBA is present.
US liquid base; residual activity after 8 weeks storage at 30°C
Effect of 4-FPBA decreases with increasing pH

14. Enzyme applications not approved due to safety considerations
4/15/2017
NOVOZYMES PRESENTATION
Enzyme applications not approved due to safety considerations .
Carpet cleaners and prespotters - household and institutional
Hard surface cleaners using high pressure delivery system - may generate breathable aerosols
Laundry prespotters using trigger sprays that deliver aerosol particles below 50 micron hence can be breathed in

15. Enzymes have multifunctional properties
4/15/2017
NOVOZYMES PRESENTATION
Enzymes have multifunctional properties .
Boost the general cleaning performance
Stain removal of organic origin (detergency)
Soil release (particulate soil removal)
Whitening (prevent soil redeposition, help with film removal)
Color Brightening (color care)
Enzymes are high performance ingredients that can replace chemicals in industrial production
Enzymes are readily biodegradable

16. In summary Enzyme offer High performance qualities
4/15/2017
NOVOZYMES PRESENTATION
In summary
Enzyme offer High performance qualities
Enzymes are Environmentally-friendly ingredients
Enzymes ensure Lower wash temperature performance