ENZYME STABILITY

IN LIQUID LAUNDRY
DETERGENTS

LUNA 2014-10619
2

CONTENT

 Enzymes for liquid laundry detergents

 Enzymatic and physical stability
 Critical parameters for enzymatic stability in
liquid laundry detergents
 Examples of mitigation plans
 Accelerated storage conditions
 Production guidelines
 Formulation guidelines
 One-pager printout
3

ENZYMES IMPROVE DETERGENCY, REMOVE

STAINS AND PROVIDE SOIL ANTI-
REDEPOSITION BENEFITS

Lipases for fat and

Cellulases for whiteness
grease stains from
and color care
oils, sebum…

Proteases for protein- Pectin-degrading enzymes

based stains such as from
blood, grass, meat.
Protease Evity and Ultra
comes with built-in
stabilization system
for fruit and pectin stains
from tomatoes, bananas,
yogurts, smoothies…

Amylases for starch-based Mannanases for mannan

stains from pasta, rice, stains from chocolate,
potatoes, sauces… thickeners, shampoo…
4
PRESERVING THE WASH PERFORMANCE
AND CHARACTERISTICS FOR A LIQUID DETERGENT
Enzymatic stability:
 Preservation of enzymatic activity
and enzyme performance
 Residual activity is enzymatic activity
after a given time compared to a
reference (frozen sample, or start
activity)
 Often accelerated conditions are
used to predict long term stability at
ambient temperature: Ex 8 weeks at
30°C.
Physical stability:
 Preservation of physical characteristic
of the detergent and basic wash
performance.
 High expertise of soapers, in rare
cases it can be impacted by enzyme
addition
 E.g. turbidity, color, pH, viscosity,
surfactant stability, phase stability,
etc…
5

ENZYMATICAL STABILITY
IN LIQUID DETERGENTS

 Enzymes are biological catalysts in the form of proteins

which have a certain 3D structure, crucial for their activity
and wash performance. Denaturation (unfolding) may
result in loss of enzyme activity and wash performance.

 Liquid detergents contain potential protein denaturants

such as surfactants and builders in direct contact with
added enzymes. However Novozymes enzymes are
developed to be stabilized towards surfactants.

 Proteases which degrade proteinous stains may also

degrade enzymes in the detergent (autoproteolysis/
proteolysis) and this is one of the most important factors
to control during storage by using protease inhibitor
systems.

 Other factors, such as the type of surfactants, pH, and

elevated temperatures can accelerate the denaturation
process.

 Accelerated conditions are often used for faster long term

predictions. Typically 8w 30C can be used.
6

CRITICAL PARAMETERS
ENZYME STABILITY IN
LIQUID DETERGENTS

• Surfactant system

• Protease inhibitor systems

• Other enzyme stabilizers

• pH

• Water activity

• Minors, ex chelators

These factors interact !

The detergent system must be
seen as a complex matrix
 7

CRITICAL PARAMETERS
SURFACTANT SYSTEM
- TYPE OF ANIONIC SURFACTANT

H3 C Different charged surfactants gives different enzyme

stabilities, the general trend is:
CH

AES (SLES) > MES > AOS ≥ LAS > Cationics

3

LAS
HC
3

alkyl ethoxy sulfate methyl ether sulfate Alpha Olefin Sulfonate linear alkylbenzene sulphonate cationic
SO
3 Na

SLES Best worst

/AES O

Substituting part of LAS with SLES increases stability.

O
O S
O
3 Na

H3C O

OMe
Nonionic surfactants reduce the negative effect of anionic
MES SO3 Na
surfactants by lowering CMC. Pure NI is generally very good
for stability.
AEO/
The ratio between AI and NI is important for stability. The more
Alcohol ethoxylate NI, the better. AI/NI preferably < 2/1 for mixed AI systems.
At high water content or at elevated pH the negative effect of
charged surfactants is higher.
8 2/11/2015 NOVOZYMES PRESENTATION – LUNA 2012-01537

CHANGING THE ANIONIC SURFACTANT

CAN INCREASE STABILITY

Protease stability in a liquid model

EU medium tier detergent with different surfactants:

2W 35C RESIDUAL ACTIVITIES

Protease: SLES > LAS/SLES > MES > LAS > AOS OF SAVINASE ULTRA XL
RELATIVE TO FROZEN
Lipase : SLES > MES > AOS >> LAS or LAS/SLES REFERENCE
Amylases: less dependent on surfactant type COMPOSITION: 12% ANIONIC; 9.5%
NONIONIC; 10% FATTY ACIDS; 5 %
MPG; BUILDERS (CITRATE+
best for stability worst for stability PHOSPHONATE). 50% WATER; PH 8.4
10

CRITICAL PARAMETERS
PROTEASE INHIBITOR SYSTEMS

Controlling (auto)proteolysis is very important for securing

stability of multi enzyme and single protease detergents.

Protease inhibitors must inhibit the protease during storage

but release quickly during wash.

The most commonly used protease inhibitor systems

• Novozymes Evity ® liquid proteases; best in class in-build
stabilization system for liquid detergents.
• Novozymes Ultra ® liquid proteases; also with in-build
stabilization system.
• Boric acid/borax+polyol (e.g. 2% of boric acid and 2%
polyol (often MPG or glycerol))
• Formate w/wo polyol (e.g. 1-2% Na-formate); very good
to boost effect of particularly Evity and Ultra proteases.

At higher pH and higher water contents, the need for protease

inhibitors is higher.
11

EVOLVING LIQUID ENZYME TECHNOLOGY

LIQUID EVITY PROTEASE

Novozymes liquid protease Evity® product range includes a

very efficient protease inhibitor delivering best in class
protease- and multi enzyme stability yielding better
preserved performance after storage:

• Increased protease wash performance after storage

• Increased multi-enzyme wash performance after storage.
Allows inclusion of more enzymes besides protease,
enabling further claim building
• Greater formulation flexibility;
replacement of boric acid in the 8 weeks
liquid detergent. Optimize your 8 weeks
Corresponding
Novozymes
formulation and explore Standard Evity® protease
Novozymes
cost savings related Unwashed
liquid protease

to capacity, storage
and transportation.

CFT PC03 (chocolate/milk/soot)

Washing conditions: EU front loader, 40°C,
15°dH, detergent dosage:
75 g/wash, EU liquid laundry detergent; samples
stored at 30°C for 8 weeks
12

EVOLVING LIQUID ENZYME TECHNOLOGY

LIQUID EVITY PROTEASE

Strong protease and multi enzyme stability yields better

preserved wash performance after accelerated storage
(8w 30°C).

Protease Multienzyme
On EMPA 117 On 9 A.I.S.E. stains
13

CRITICAL PARAMETERS
OTHER ENZYME STABILIZERS

Other enzyme stabilizers include:

• Calcium:
• Ca2+ is important for stability of proteases and amylases.
• Its recommended to use 0.02-0.06w% CaCl2.
• Strong builders should be used with caution and checked
for compatibility.
• For detergents with high fatty acids/soaps; high overall
surfactant levels; and/or HEDP, CaCl2 may cause
precipitations; and lower CaCl2 levels may be needed.
• Polyols:
• preferentially excluded excipients/solvents, such as
polyols (e.g. glycerol, propylene glycol/MPG, dipropylene
glycol/DPG, Sorbitol, etc.) stabilize enzymes. Use 2-5%
• Keep Ethanol below 5%.
• Salts; Ex Na-formate
• lower water activity and can increase the effect of
protease inhibitors, particularly 0.5-2% Na-formate is
itself a weak protease inhibitor and boosts the effect of
Evity ® and Ultra® protease inhibitors.
14

CRITICAL PARAMETERS
pH OF THE DETERGENT

High pH increases the risk of enzyme degradation

At high pH proteases are more active and negative effect of

surfactants is higher and more protease inhibitors are needed.

pH 6.5-8.5 pH 8.5-9.5: pH 9.5-11:

Advisable. In multi-enzyme Possible. Will Generally not
cocktails; protease inhibitors require protease advisable.
inhibitors and
and/or enzyme stabilizers may
enzyme stabilizers
be needed for securing second
enzyme stability.

Check wash pH: When the recommended detergent dose is

low, pH of the wash water is less impacted by detergent pH,
and then pH of the detergent can beneficially be lowered
without detrimental effect on wash performance.
 15 2/11/2015 NOVOZYMES PRESENTATION – LUNA 2012-01537

EFFECT OF pH ON
PROTEASE STABILITY

Protease stability.
Preferred pH 7-9

Residual activity

100%
4w37C
80%

60%

40%

20%

0% pH 4W 37C RESIDUAL ACTIVITIES

7.5 8.0 8.5 9.0 9.5 10.0 RELATIVE TO FROZEN REFERENCE
COMMERCIAL EU LIQUID DETERGENT
WITH 1% SAVINASE ULTRA XL.
PH ADJUSTED WITH NAOH/HCL
16

CRITICAL PARAMETERS
WATER CONTENT

Protease stability and multi enzyme stability with protease is

highly depending on water content.

At high water content, proteolytic activity is higher and more

protease inhibitor levels or more efficient inhibitors are needed.

At high water content negative effects of e.g. LAS; high pH are

stronger.

Lowering the water content generally stabilize enzymes and

reduces proteolytic activity and need for protease inhibitors

Water content can be lowered by:

• Compaction (increasing surfactant/reducing water)
• Increasing polyol content or adding polymers/salts
• Liquid unit doses have low water content
17

CRITICAL PARAMETERS
MINORS

“Minors” are detergent ingredients which are not surfactants,

water, enzymes and enzyme stabilizers

Most preservatives are OK Formaldehyde is not

(isothiazolinones ex BIT, MIT, CMIT).
Mild to moderate builders are
generally OK. Ex most phosphonates
(DTMPA, EDTMPA) and organic acids (ex
citrate) with a certain threshold level
securing some free Ca in the detergent.
Strong builders should be used with
caution (ex HEDP; DTPA) but can in
many cases be incorporated.
Perfumes, colorants, opacifiers,
optical brightners, polymers are
generally OK.
compatible with enzymes.
Formaldehyde releasing
preservatives to be used with
caution.
EDTA is not compatible with
most enzymes in liquid
laundry detergents.
Bleaches (peroxide and
chlorine) are generally not
compatible with enzymes in
liquid detergents.
18 2/11/2015 NOVOZYMES PRESENTATION

CHELATORS / BUILDERS / SEQUESTRIANTS

COMPATIBILITY WITH ENZYMES

Mild to moderate builders are generally OK.

Strong builders should be used with caution but can

in many cases be incorporated.

Generally OK DTMPA Citrate

EDTMPA DEG

Recommended to test; DTPA GLDA

but in many cases HEDP IDS
possible EDDS MGDA
Not recommended or at EDTA
least thouroughly tested
ENZYME COMPATIBILITY WITH
CHELATORS ALSO DEPEND ON
Enzyme compatibility in the presence of chelators can AMOUNT OF CHELATOR; THE
be improved by adding small amounts of Ca to the REST OF THE DETERGENT
COMPOSITION; AND ON
detergent formulation if allowed for physical stability ENZYME CLASS
reasons.
19

MITIGATION PLAN
HANDLING CRITICAL PARAMETERS BY
FORMULATION OPTIMIZATION.

Examples Mitigations for optimal enzyme stability

High water content Use efficient protease inhibitor (Evity® / Ultra®)
and/or low possibly combine with 1-2% Na-formate.
surfactant Use 0.02-0.06% CaCl2.
concentration Reduce pH to 7-8’ish.
Use mixed surfactant systems (LAS/SLES);
preferably with AI/NI ratio 1/1 or even better 1/2.
High AI/NI ratio Use efficient protease inhibitor (Evity® / Ultra®)
needed for possibly combine with 1-2% Na-formate.
performance or Use 0.02-0.06% CaCl2.
cost Reduce pH to 7-8’ish.
Use mixed AI systems (LAS/SLES).
Add 2-5% polyol
Reducing Enzyme cocktail added for improved performance.
surfactant Enzyme stabilizers added to manage the resulting
concentration higher water content.
without Use efficient protease inhibitor (Evity® / Ultra®)
compacting
20 2/11/2015 NOVOZYMES PRESENTATION

EXAMPLE OF MITIGATION
LOWERING pH AND CHANGING ANIONIC TYPE
FROM LAS TO SLES INCREASES STABILITY

Protease stability as a function of pH and AI type:

Original pH 8,75 and pure LAS () resulting in

compromized protease stability
Optimized formulation; pH 8.0 and LAS:SLES 1:1 ()
and highly improved protease stability and broader
cleaning profile.
4W 30C RESIDUAL ACTIVITIES
RELATIVE TO FROZEN REFERENCE
SAVINASE ULTRA® 16XL
COMPOSITION 12% ANIONIC (LAS/SLES);
9.5% NONIONIC; 10% FATTY ACIDS; 5%MPG;
BUILDERS (PHOSPHONATE+CITRATE);
APPROX 50% WATER.
21 2/11/2015 NOVOZYMES PRESENTATION

EXAMPLE OF MITIGATION
IMPROVING STABILITY AFTER REDUCTION OF
SURFACTANT LEVEL, MAINTAINING WASH PERFORMANCE

Original:
Surfactant level ~30%
pH 8.4

Surfactant reduced:
Surfactant level ~ 20%
included in original
pH 8.0
Stabilizers:
+0,02% CaCl2
XPect not

+3% glycerol+3.5% MPG

+2% Na-formate

RESIDUAL ACTIVITIES
RELATIVE TO FROZEN REFERENCE.
EU MODEL DETERGENT WITH
MULTIENZYME COCKTAIL (SAVINASE
ULTRA® XL; LIPEX; XPECT; STAINZYME)
Composition original:
12% LAS; 9.5% nonionic; 10% fatty acids;
5% MPG; Builders; approx 50% water.
22

CHOOSING ACCELERATED CONDITIONS

THAT PREDICT LONG TERM STABILITY
Warehouse Study
in China.
Guangzhou; July
and August. Highest
temp was 32C and
lowest was 28C.
Average temp was
30C

Truck transport
Study in China. End Consumer
Tianjin – Guangzhou Study world wide.
(2200Km, 2 days), July Half of the shelf life
and August. Temp in is at customer. 13
products did not countries in their
fluctuate on outdoor respective summer
temp, and temp was in times. Average
average 30C temp in detergent
box did not exceed
30C

 In warm places on earth; average temp is around 30°C and in most part of
the world; 30°C would be accelerated conditions
 To be best prepared for the journey, we recommend choosing the best in class
stabilized enzymes, like Novozymes Evity.
23 2/11/2015 NOVOZYMES PRESENTATION

USING ACCELERATED CONDITIONS TO

PREDICT LONG TERM STABILITY

Typically 8w 30°C is a good accelerated condition for predicting

long term stability at ambient temperature.

Increasing temperature may lead to less

predictive results. RESIDUAL ACTIVITIES
RELATIVE TO FROZEN REFERENCE
Rate of decay depend on specific enzyme type and on the EU COMMERICAL DETERGENT WITH
detergent composition and mechanism of decay. MULTIENZYME COCKTAIL
(INCL PROTEASE)
25

PHYSICAL STABILITY
OF LIQUID DETERGENTS
 Physical stability concerns preservation of physical characteristic of the
detergent and basic wash performance. Ex clarity; phase stability; intactness
of other functional ingredients.

 When testing phase stability at high temperatures (50 or 60°C); its

recommended to exclude enzyme as they may be partly heat inactivated and
hence influence the result.

 The physical stability of a liquid detergent can in rare cases be affected by

enzyme addition, and may be related to:
 the enzyme itself (e.g. haze) mostly related to low solubility in low water or
non-polar detergents or in combination with compromised stability
 Ca-precipitations (or haze) if the detergent has too little Ca-tolerance ex due
to high HEDP or high fatty acid (All liquid enzymes contains small amounts of Ca);
need 50-100 ppm Ca-tolerance
 stabilizers (salts, ex Na-formate; polyols) added for enzyme stabilization (haze,
viscosity or phase change)
 Related to enzyme activity (very rare), ex lipase and ester based ingredients.

 Other issues are rarely related to the enzyme addition.

26

PRODUCTION GUIDELINES
ADDITION OF
ENZYMES

When adding liquid enzymes to detergents:

• Add in the end of the process (ex like colors and/or

perfumes) where the temperature after
neutralization/solubilization is low again (<40°C-ish),
dissolution steps are completed and pH is stable.
• Preferably add enzymes in-line separately (single
enzyme or pre-manufactured liquid blends like Medley®).
For batch production; add one at a time (no specific
order is needed) and secure good stirring/mixing
(ex 15-30 min depending on viscosity).

• Avoid dilution of enzyme products, but if needed, use

polyol/water mixtures. Its not recommended to blend
separate enzymes before mixing into detergents;
instead use Novozymes Medley® line.
• Its not recommended to premix enzymes with other
ingredients prior to addition to detergent, like ex colors,
perfumes, preservatives.
27

PRODUCTION GUIDELINES
ADDITION OF ENZYME
STABILIZERS

When adding enzyme stabilizers to the detergent:

• Calcium; boric acid/borax and Na-formate can be

added at any time as long as full dissolution is
achieved.
• For concentrated detergents Na-formate and/or boric
acid/borax can beneficially be added before or during
heating step (neutralization and/or dissolution of
surfactants). Na-formate/formic acid and boric acid
may change pH and require more/less neutralization
chemicals.
• Polyols can be added at any time; as solvent initially;
as viscosity controller in the end; or to be used as
premixing/dilution of separate enzymes in the end of
the process.
• Ca2+ can beneficially be added as a solution, and the
physical stability of the detergent should be checked.
FORMULATION
GUIDELINES LIQUID
LAUNDRY DETERGENTS
29

SURFACTANT SYSTEM

Charged surfactants are negative for enzyme stability. Nonionic

surfactants reduce the negative effect of anionic surfactants.

Combine NI and AI and have AI/NI < 2/1.

Best for stability worst for stability

AI/NI: 0/1 >> 1/2 > 1/1 > 2/1 >> 1/0
AI: AES > MES > AOS = soap > LAS
pure AES > LAS/AES > pure LAS
NI: All NI are positive for stability
Long ethoxylations are good for solubility
Counter ion Neutralization with MEA or KOH gives highest
surfactant solubility
Total amount of surfactants 15-70% is possible; in the low end
we need protease inhibitors and lower AI/NI ratios.
30

PROTEASE INHIBITOR
SYSTEMS

Protease inhibitors are needed to decrease (auto)proteolysis in

most detergents containing protease.

More efficient protease inhibitors are needed when

• pH of the detergent is high
• Water content is high
• The surfactant system is high in AI.

The most commonly used protease inhibitor systems

• Novozymes Evity ® liquid proteases; best in class in-build
stabilization system for liquid detergents.
• Novozymes Ultra ® liquid proteases; also with in-build
stabilization system.
• Boric acid/borax+polyol (e.g. 2% of boric acid and 2%
polyol (often MPG or glycerol))
• Formate w/wo polyol (e.g. 1-2% Na-formate); very good
to boost effect of particularly Evity and Ultra proteases.
31

OTHER ENZYME
STABILIZERS

• Calcium: Ca2+ is important for stability of proteases and

amylases. Use 0.02-0.06 w% CaCl2 but check compatibility
with strong builders and fatty acids. All enzyme
formulations contain small amounts of Ca. The detergent
must have Ca-tolerance to deal with 50-100 ppm Ca.

• Polyols stabilize enzymes, e.g. glycerol, propylene

glycol/MPG, dipropylene glycol/DPG, Sorbitol, etc. Use 2-
5%. Keep Ethanol below 5%.

• Salts can increase the effect of protease inhibitors,

particularly 0.5-2% Na-formate is boosting the effect of
Evity ® and Ultra® protease inhibitors.
32

DETERGENT pH
AND WATER CONTENT

At high pH and high water content/activity; proteolysis is

higher, and more protease inhibitors and enzyme stabilizers
are needed. Likewise, the negative effect of charged
surfactants is higher.
pH Water Recommendation
in multi enzyme solutions; protease
inhibitors like Evity ® or Ultra ® and/or
6.5- 5-50
Advisable stabilizers are needed. In low water
8.5 w%
formulations (unit doses) need for
protease inhibitors is lower
Will require efficient protease inhibitors
50-80
like Evity and enzyme stabilizers, possibly
8.5- w%
combined with Ca and Na-formate.
Possible 9.5
Will require efficient protease inhibitors
>80
combined with Ca and Na-formate and
w%
enzyme friendly AI/NI ratio
Not 9.5- >85 pH can in some cases be lowered and
advisable 12 w% performance regained by enzymes.
33

MINORS

“Minors” are detergent ingredients which are not surfactants,

water, enzymes and enzyme stabilizers

Most preservatives are OK Formaldehyde is not

(isothiazolinones ex BIT, MIT, CMIT).
Mild to moderate builders are
generally OK. Ex most phosphonates
(DTMPA, EDTMPA) and organic acids (ex
citrate) with a certain threshold level
securing some free Ca in the detergent.
Strong builders should be used with
caution (ex HEDP; DTPA) but can in
many cases be incorporated.
Perfumes, colorants, opacifiers,
optical brightners, polymers are
generally OK.
compatible with enzymes.
Formaldehyde releasing
preservatives to be used with
caution.
EDTA is not compatible with
most enzymes in liquid
laundry detergents.
Bleaches (peroxide and
chlorine) are generally not
compatible with enzymes in
liquid detergents.
34

PHYSICAL
STABILITY

Physical stability of a detergent can in rare cases be influenced

by enzyme addition or by inclusion of enzyme stabilizers.

The detergent should preferable have possibility for inclusion of

0.02% CaCl2 both to give better stability and to deal with small
amounts of Calcium coming from enzyme formulations.

Enzyme stabilizers like Na-formate may slightly change

visual appearance in clear concentrated detergents; try adding
during heating step or reduce amount to 0.5-1%.

Enzymes can have too low solubility in non-polar and/or low

water detergents (e.g. unit doses with high enzyme inclusions).
• Increase polarity of the detergent; add e.g. salts; water;
glycerol/MPG (~2%); hydrotrope (~2%); EtOH (~2%).
• Change ratio between AI/NI to more AI
• change pH

Avoid some ester based ingredients when using Lipase

35
PRODUCTION
GUIDELINES
Start, Mixing; solubilization
solvent neutralization
Enzyme stabilizers such as
CaCl2; boric acid/borax and
Na-formate can be added at
any time as long as full
dissolution is achieved.
For concentrated detergents; it
beneficial to add them during
heating / neutralization step.
Polyols can be added at any
time.
Finetuning and
standardization
Add enzymes in the end of
the process (< 40°C and
final pH). In-line
separately or if batchwise,
in separate steps with in-
between mixing.
If its needed to dilute the
enzyme liquid products;
use polyol/water mixtures
36

SUMMARY

Surfactant Water content (w%) Detergent pH and

concentration Water activity WA Protease inhibitor system
>60%. <10w%
Generally ok, but solubility might pH outside 7-9
QUESTION-
Careful about possible solubility be an issue. Depend much on the too low inhibitor levels or no
ABLE
issues in non-polar detergents solvent system. Protease inhibitors protease inhibitors at all
is not necessarily needed.
40-60%
pH 6,5-8,5:
Use mixed surfactant systems. 10-50w% Protease inhibitors like Evity
Protease inhibitors like Evity ® or ® or Ultra ® or Boric acid;
Preferably AI:NI < 2:1.
Ultra ® and polyols might be Ca and polyols might be
High polyol and MEA might be needed needed
ADVISABLE needed for surfactant solubility.
pH 8,5-9,5:
60-80w%
20-40% Protease inhibitors like Evity
Protease inhibitors like Evity ® or
® or Ultra ® combined with
Use mixed surfactant systems. Ultra ® possibly combined with Na-
Ca , polyol and Na-formate
Preferably AI:NI < 2:1. formate and Ca. pH preferably 7-
might be needed
8.5.
80-85w%
15-25% Protease inhibitors like Evity ® or pH outside 7-9
QUESTION- Ultra ® combined with Ca , and Na- too low inhibitor levels or no
ABLE Use mixed surfactant systems. formate is needed. combination with Na-formate
Preferably AI:NI < 1:1. Use higher protease doses (>0.2%
Evity L/XL)
>85w%
NOT unless very favourable surfactant
Pure AI system or pure LAS pH > 9,5
ADVISABLE composition and high inhibitor
levels.
37

FORMULATING LIQUID DETERGENTS

FOR OPTIMAL ENZYME STABILITY
POLYOLS: SURFACTANT SYSTEM:
Use min 2-5% polyol; Use mixed surfactant systems
ex MPG or glycerol. keeping anionic : nonionic ratio 1:1
or even better 1:2. Preferably
Particularly with combine LAS and SLES.
Ultra® or boric acid
protease stabilization

CALCIUM:
Use 0,02-0,06% CaCl2 PROTEASE
for optimal protease INHIBITORS:
and amylase stability
Use protease inhibitor
systems. Ex
Novozymes protease
Evity® or Ultra ®.

FORMULATION pH
Preferably have pH 6.5-8.5 and
combine with protease inhibitor
systems for multi-enzyme
cocktails.
CONSULT
your Novozymes
technical contact for
MINORS further questions or
assistance
ORDER OF ADDITION Most “minors” are OK.
Preferably add CaCl2 and Na-formate during Strong chelators should be used with caution.
heating step and add enzymes individually at Avoid bleach and formaldehyde releasing
the end when temperature is again low. preservatives.