Enzymes used in the detergent industry

Enzymes used in the detergent industry
After the introduction to Detergents we now turn the focus to enzymes that are a used in combination with surfactants in many applications in households, institutions and industries. Please let me introduce myself: I am chemical engineer. My background within enzymes dates 9 years back. I have worked with enzyme applications within detergent, textile and the forest products industry. I am currently responsible for technical service for the detergent industry in the Nordic countries, Benelux, France and Italy. My special areas are Cellulases and Amylases. In the following I will go through the basis for use of enzymes in the detergent area. 13/04/2017 NOVOZYMES PRESENTATION NOVOZYMES A/S

4 Types of enzymes are used for different detergents
13/04/2017 NOVOZYMES PRESENTATION 4 Types of enzymes are used for different detergents Substrate Examples Blood, egg, milk grass, meat Protease Proteins Organic fats and oils, sebum, lipstick Lipase Triglycerides Proteases and Lipases are specifically hydrolysing proteins and fats respectively. A given protease or lipase is able to hydrolyse a lot of different proteins or fats. Carbohydrases are evenmore specific: They hydrolyse only one type carbohydrate. Both starch and cellulose contains linear polymers of glucose. To be able to hydrolyse both you need two different enzymes, because strarch contains alfa bonds and cellulose beta-bonds. This is why detergents may contain only one protease, one lipase but several carbohydrases. We know of products with 4 different carbohydrases. In the following I will explain how the enzymes work. Amylase Starch Rice, noodles, potatoes, corn Carbohydrases Cellulase Cellulose Cotton, viscose NOVOZYMES PRESENTATION NOVOZYMES A/S

Catalytic actions of detergent enzymes
13/04/2017 NOVOZYMES PRESENTATION Catalytic actions of detergent enzymes

13/04/2017 NOVOZYMES PRESENTATION Stain removal Surfactants can remove most of the soils in laundry. Why do we need the enzymes? Enzymes and surfactants generally work complementary, and together they can do the cleaning job at low temperature + mild pH The following cartoon shows how enzymes degrade soil partially and surfactants bring the reminder into suspension

Removal of stain 1: The stain is bound to a surface
13/04/2017 NOVOZYMES PRESENTATION Removal of stain 1: The stain is bound to a surface Stain Substrate Fibre The slide shows a schematic drawing of a soil particle containing protein on a textile fibre. The protein is imagined to glue the soil to the fibre. NOVOZYMES PRESENTATION NOVOZYMES A/S

Removal of stain 2: A surfactant has been added
13/04/2017 NOVOZYMES PRESENTATION Removal of stain 2: A surfactant has been added Soil Fibre Surfactant Surfactants (”the red microphones”) will adsorb with the hydrophobic ends to the soil, so the the hydrophilic end points out into the water. The soil however resists going into solution due to the protein glue. NOVOZYMES PRESENTATION NOVOZYMES A/S

Removal of stain 3: Enzymes are added and degrade available sites
13/04/2017 NOVOZYMES PRESENTATION Removal of stain 3: Enzymes are added and degrade available sites Soil Fibre Surfactant Enzyme With addition of protease (The green UFO’s) the protein acting as glue is hydrolysed into small soluble peptides NOVOZYMES PRESENTATION NOVOZYMES A/S

Removal of Stain 4: The soil comes loose and goes into suspension
13/04/2017 NOVOZYMES PRESENTATION Removal of Stain 4: The soil comes loose and goes into suspension Surfactant Enzyme Soil When the protein is gone the surfactant can take the soil into solution. As catalysts proteases are not used up by the reaction why there are still some around. Fibre NOVOZYMES PRESENTATION NOVOZYMES A/S

General principle of stain removal
13/04/2017 NOVOZYMES PRESENTATION General principle of stain removal Clean vest Stain Wash, no enzýmes Stain less visible  Wash with enzymes Stain no longer visible Illustration of the test method. Starch spots (without visible soil) are applied. The fabric is washed several times. The starch spots cannot been seen - but they are still there. In the next step the fabric is exposed to a soil suspension - without detergent. This makes the spots visible because the starch binds the particulate soil. The fabrics are then washed again in detergent. The visible soiling is only removed slowly. After some wash cycles it is still visible. This can be continued with more soiling-washing cycles. The original starch stays on the fabric even after many detergent wash cycles, if there is no amylase present.  NOVOZYMES PRESENTATION NOVOZYMES A/S

Removal of invisible stains: redeposition
13/04/2017 NOVOZYMES PRESENTATION Removal of invisible stains: redeposition Clean vest Stain Washed  Washed in dirty water, No enzymes Washed in dirty water, With enzymes Illustration of the test method. Starch spots (without visible soil) are applied. The fabric is washed several times. The starch spots cannot been seen - but they are still there. In the next step the fabric is exposed to a soil suspension - without detergent. This makes the spots visible because the starch binds the particulate soil. The fabrics are then washed again in detergent. The visible soiling is only removed slowly. After some wash cycles it is still visible. This can be continued with more soiling-washing cycles. The original starch stays on the fabric even after many detergent wash cycles, if there is no amylase present.   Conclusion: Enzymes give more permanent cleaning! NOVOZYMES PRESENTATION NOVOZYMES A/S

Removal of inorganic particulate soils
13/04/2017 NOVOZYMES PRESENTATION Removal of inorganic particulate soils H1: washed without cellulase H2: washed with cleaning cellulase

Removal of fuzz and pills
13/04/2017 NOVOZYMES PRESENTATION Removal of fuzz and pills Without cellulase With the care cellulase: Carezyme®

Enzymes boost the removal of …
13/04/2017 NOVOZYMES PRESENTATION Enzymes boost the removal of … Food and plant stains in laundry and ADW Invisible impurities in laundry and ADW Particulate inorganic soils in laundry Fuzz and pills in laundry BUT….. Which enzymes for what?

International brand support model
13/04/2017 NOVOZYMES PRESENTATION International brand support model Cellulase Lifestyle Aesthetics Lipase Amylase Enzymes help you renew your brand and drive consumer loyalty (the purchase – trial –re-purchase cycle). They are a strong supporter of this process and strong tool in supporting your brand. Protease Basic performance NOVOZYMES PRESENTATION NOVOZYMES A/S

International brand support model
13/04/2017 NOVOZYMES PRESENTATION International brand support model Polarzyme Stainzyme

NOVOZYMES PRESENTATION
13/04/2017 NOVOZYMES PRESENTATION Proteases Proteases, the slide shows a ball molecule model of Savinase where Carbon is green, Oxygen red and Nitrogen blue. The protease is a so called globular protease. Proteases are the most important of enzyme for laundry detergents, wheras in dishwash detergents it is secondary to amylases. In laundry detergents proteases are mainly used for stain removal. In the following seriess of slides the pretease action is described further. Protease -R1CH-CO-NH-CR2H- + H2O « -CR1H-COO- + +H3N-CR2H- NOVOZYMES PRESENTATION NOVOZYMES A/S

Proteases from Novozymes
13/04/2017 NOVOZYMES PRESENTATION Proteases from Novozymes Alcalase® For low pH, soft on wool Esperase® For high pH, mainly I&I* Everlase® Relatively stable in presence of bleach Savinase® High performance standard protease Polarzyme® High performance low temp. protease Ovozyme® Tolerates protease inhibitors from eggs Relase® For liquid detergents Coronase® For liquid detergents *I&I = Industrial and Institutional cleaning

Proteases catalyze the hydrolytic cleavage of the peptide chain
13/04/2017 NOVOZYMES PRESENTATION Proteases catalyze the hydrolytic cleavage of the peptide chain Protease -R1CH-CO-NH-CR2H- + H2O → -CR1H-COO- + +H3N-CR2H- Proteases can provide: stain removal cleanliness general whitening Proteases are the most widely used enzymes. In laundry detergents, protein stains such as grass, blood, egg and human sweat are removed through proteolysis. In automatic dishwashing detergents, proteases secure the removal of proteinaceous food films, which are a particular problem with glassware and cutlery. Proteases are classified according to their source of origin (animal, plant, microbial), their catalytic action (endo-peptidase or exo-peptidase) and the nature of the catalytic site (active site). Common names and trade names, typical pH ranges and preferential specificity are used to characterize them. Based on a comparison of active sites, catalytic residues, and three-dimensional structures, four major protease families are recognized: serine, thiol, aspartic and metalloproteases. The serine protease family contains two sub-groups: chymotrypsin-like and subtilisin-like. The latter is the most important group for detergent applications. NOVOZYMES PRESENTATION NOVOZYMES A/S

Facts about proteases Novozymes proteases have been used since 1963
13/04/2017 NOVOZYMES PRESENTATION Facts about proteases Novozymes proteases have been used since 1963 Are used in 75 % of all detergents Protease is a basic ingredient in laundry and ADW detergents Protease effects cannot be substituted by any other ingredients

Protease effect on blood spots
13/04/2017 NOVOZYMES PRESENTATION Protease effect on blood spots LHjo00070 and LHjo00103, blood spots on B T-shirts 1ml blood applied, dried overnight before washing Scanner settings 95, 135 Scan files , , Full-scale wash, A/P conditions, 12 minutes, 25°C Detergent: Surf, Hindustan Lever, 1,5g/l Savinase 12T, 0,5% not washed washed 3 times, no protease washed 3 times, with protease NOVOZYMES PRESENTATION NOVOZYMES A/S

Stain removal with protease in laundry
13/04/2017 NOVOZYMES PRESENTATION Stain removal with protease in laundry Protease no yes Protease no yes Cocoa (CFT) Blood (CFT) Salmon paste (Equest) Grass (Equest) Spinach (Equest) Olives (Equest)

13/04/2017 NOVOZYMES PRESENTATION Protease effect on blood spots Protease effect after 3 wash cycles and re-soiling No protease Re-soiling With Protease LHjo00070, blood spots on B T-shirts 1ml blood applied, dried overnight before washing Scanner settings 95, 135 Files , -02, -03, -04 Full-scale wash, A/P conditions, 12 minutes, 25°C Detergent: Surf, Hindustan Lever, 1,5g/l Savinase 12T, 0,5% Resoiling treatment: Merck carbon, cat 2186, 1,0g in 42l water, wash 3 minutes and rinse. Re-soiling NOVOZYMES PRESENTATION NOVOZYMES A/S

Proteases perform well at high pH
13/04/2017 NOVOZYMES PRESENTATION Proteases perform well at high pH The figure shows Alcalase versus Savinase Relative Activity (%) pH

Wash performance of polarzyme 12T
13/04/2017 NOVOZYMES PRESENTATION Wash performance of polarzyme 12T 25 °C, TOM1, Regular powder, 10 °dH, 20 min, EMPA 117 swatch Unwashed Light reflection Dose-response plateau 1) TOM (Tergotometer) is a small scale method simulating top loading US washing machine NOVOZYMES PRESENTATION NOVOZYMES A/S

Protease sensitive test swatches:
13/04/2017 NOVOZYMES PRESENTATION Protease sensitive test swatches: CFT C-10 Pigment/Oil/Milk on cotton CFT C-03 Chocolate milk/Soot on cotton EMPA112 Cocoa on cotton CFT C-05 Blood/Milk/Ink on cotton EMPA116 Blood/Milk/Ink on cotton EMPA117 Blood/Milk/Ink on cotton/polyester EMPA111 Blood on cotton CS Grass on cotton PCS Grass on cotton/polyester EMPA Grass on cotton WfK 10N Egg on cotton WfK 20N Egg on cotton/polyester Light reflection Dose-response plateau NOVOZYMES PRESENTATION NOVOZYMES A/S

Low temperature profile of polarzyme
13/04/2017 NOVOZYMES PRESENTATION Low temperature profile of polarzyme Activity profile of Polarzyme shifted towards lower temperatures maintaining activity at high temperature Activation temperature: around 70C. Polarzyme a true low-temperature protease NOVOZYMES PRESENTATION NOVOZYMES A/S

Protease benefit tested in wash & wear test
13/04/2017 NOVOZYMES PRESENTATION Protease benefit tested in wash & wear test Consumer wash and wear test: 70 men asked to wear a white cotton T-shirt T-shirts collected and cut in half One half washed in detergent with 0.1% Polarzyme 12T Other half washed in well known non-biological brand T-shirts sewed up, redistributed for another wearing and washing cycle (4 cycles in total) After each cycle the person responsible for the family´s laundry was asked to rate the result

Protease benefit in wash & wear test - overall consumer preference
13/04/2017 NOVOZYMES PRESENTATION Protease benefit in wash & wear test - overall consumer preference

Protease benefit in wash & wear test
13/04/2017 NOVOZYMES PRESENTATION Protease benefit in wash & wear test consumer preference for each performance attribute

13/04/2017 NOVOZYMES PRESENTATION Status for proteases Protease is applied in laundry and ADW for complete removal of protein Protease is recognised world-wide as a neccesary provider of basic detergency

13/04/2017 NOVOZYMES PRESENTATION Lipases serine-histidin-aspartic acid triade. Active site is located in a hydrofobic binding pocket, where a lid covers the active site. The lid is closed in aqupous conditions and the lid opens when the lipase meet oil/water interfases. the lid opens and the active site becomes accessible. NOVOZYMES PRESENTATION NOVOZYMES A/S

+ + Lipase mechanism Lipase No odor Lipase Odor
13/04/2017 NOVOZYMES PRESENTATION Lipase mechanism Lipase No odor + Lipase Odor + Short chain fatty acids (C4 –C8) Dairy soils

Lipases from Novozymes
13/04/2017 NOVOZYMES PRESENTATION Lipases from Novozymes Lipolase® Original lipase wild type lipase. Multi wash lipase Lipex® First wash protein engineered Lipase. Stain removal and anti redeposition effects. Lipex – PE variant of Lipolase NOVOZYMES PRESENTATION NOVOZYMES A/S

Effects with Lipase First Lipase introduced in 1988
13/04/2017 NOVOZYMES PRESENTATION Effects with Lipase First Lipase introduced in 1988 First protein engineered lipase introduced in 1998 Lipase can to some extent replace surfactants Remove fatty matter at low temperature Provide stain removal Give in-depth cleaning Reduces redeposition General whitening + Lipase - Lipase Lipstick

Activity of Lipolase during drying
13/04/2017 NOVOZYMES PRESENTATION Activity of Lipolase during drying Relative Activity (%)

13/04/2017 NOVOZYMES PRESENTATION Lipase performance Lipolase works best at low water concentration i.e. during drying of laundry Lipex also works at high water concentrations such as in the main wash

Lipase for fat removal from outside of cotton fibres
13/04/2017 NOVOZYMES PRESENTATION Lipase for fat removal from outside of cotton fibres no lipase with lipase Lard Sebum Source.: Obendorf et al., Slide prepared by: Call One cycle wash in TOM, using European conditions, without lipase and with LipoPrime. – similar to LIPEX Fibres treated with OsO4, electron microscopy Olive Oil NOVOZYMES PRESENTATION NOVOZYMES A/S

Lipase effect on inside of cotton fibres
13/04/2017 NOVOZYMES PRESENTATION Lipase effect on inside of cotton fibres Cross sectional specimen of yarns soiled with olive oil Source.: Obendorf et al., Slide prepared by: ?? Wash conditions as for Fig 8a LIPEX efect Olive oil Unwashed Olive oil Washed with detergent Olive oil Washed with detergent & lipase NOVOZYMES PRESENTATION NOVOZYMES A/S

Lipase 1’st wash effect on polyester
13/04/2017 NOVOZYMES PRESENTATION Lipase 1’st wash effect on polyester 0.2 mm washed without lipase 0.2 mm Source: Call/ThoR990292 Light microscope pictures of 100% polyester fabric, showing soil redeposition during washing, and the effect of lipase. LIPEX effect washed with lipase NOVOZYMES PRESENTATION NOVOZYMES A/S

Lipex - visual soil removal in 1st wash
13/04/2017 NOVOZYMES PRESENTATION Lipex - visual soil removal in 1st wash No Lipex Lipex 500 LU/L Lipex 1000LU/L

13/04/2017 NOVOZYMES PRESENTATION Anti-redeposition benefits after 1st wash - a secondary effect of Lipases Unwashed textiles - Lipex + Lipex One example or proof of the anti redeposition phenomenon is this trial where two large ballast swatches of cotton interlock and polyester (woven), soiled with lard/carbon black, are washed together with a lipstick swatch (wfk20LS2), and two small clean tracer swatches in TOM. When there is Lipex present we observe that the redeposition of lipid/carbon black is reduced significantly, even at such a low concentration as 300 LU/l wash solution. Especially the edges of the tracer swatches are much reduced in black color. Also the lipstick swatch are without black ”development” with carbon black and at the same time there is visual enzyme performance on this swatch. wfk20LS2 + two ballast swatches with lard/carbon black, TOM, 40C, 20 min. Heavy duty EU powder detergent (4.5 g/l). 300 LU/l NOVOZYMES PRESENTATION NOVOZYMES A/S

13/04/2017 NOVOZYMES PRESENTATION Status for Lipases Lipase can be used in laundry for complete lipid removal especially at lower temperatures Secondary effect of lipases are contribution to general cleanness by preventing redeposition of fats, oils and colorants In depth cleaning of textile fibres There is a smell issue that limits the dosage

13/04/2017 NOVOZYMES PRESENTATION Amylases This model is not really an amylase but who can tell ? NOVOZYMES PRESENTATION NOVOZYMES A/S

Amylases from Novozymes
13/04/2017 NOVOZYMES PRESENTATION Amylases from Novozymes Termamyl® Original amylase for high temp. Termamyl® Ultra Highly stable liquid amylase Duramyl® Improved stability in the presence of bleach Stainzyme® High performance “Stainfighter” Performs well under low temperature wash conditions

13/04/2017 NOVOZYMES PRESENTATION a-Amylases Amylases hydrolyse a-1,4-glucose-linkages randomly forming soluble dextrins and oligosaccharides The viscosity of the gelatinized starch is reduced and the solubility enhanced During enzymatic liquefaction, the a-1,4-linkages in starch (amylose and amylopectin) are hydrolyzed at random. This reduces the viscosity of the gelatinized starch and increases the solubility of attached starch, which is converted to water-soluble dextrins and oligosaccharides. Therefore, Termamyl, Duramyl, and BAN are often referred to as "liquefying amylases". NOVOZYMES PRESENTATION NOVOZYMES A/S

Removal of soils containing starch
13/04/2017 NOVOZYMES PRESENTATION 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

Removal of invisible starch stain
13/04/2017 NOVOZYMES PRESENTATION Removal of invisible starch stain Clean vest Starch stain Washed in any detergent Washed in dirty water, No enzymes Washed in dirty water, With enzymes Illustration of the test method. Starch spots (without visible soil) are applied. The fabric is washed several times. The starch spots cannot been seen - but they are still there. In the next step the fabric is exposed to a soil suspension - without detergent. This makes the spots visible because the starch binds the particulate soil. The fabrics are then washed again in detergent. The visible soiling is only removed slowly. After some wash cycles it is still visible. This can be continued with more soiling-washing cycles. The original starch stays on the fabric even after many detergent wash cycles, if there is no amylase present. NOVOZYMES PRESENTATION NOVOZYMES A/S

pH-profile of Termamyl and Stainzyme
13/04/2017 NOVOZYMES PRESENTATION pH-profile of Termamyl and Stainzyme

Temp. profile for Termamyl and Stainzyme
13/04/2017 NOVOZYMES PRESENTATION Temp. profile for Termamyl and Stainzyme

13/04/2017 NOVOZYMES PRESENTATION Washing time

13/04/2017 NOVOZYMES PRESENTATION Different pH profiles

Potato starch With Termamyl® With Stainzyme®
13/04/2017 NOVOZYMES PRESENTATION Potato starch Cotton CFT CS-27 Polyester CFT PS-27 Nylon CFT NS-27 Polyacryl CFT PAS-27 Cotton/Polyester CFT PCS-27 With Termamyl® With Stainzyme® Commercial powder with bleaching agent, dosage: 4 g/l, Commercial washing machine, temperature: 40°C, short programme

Rice starch With Termamyl® With Stainzyme®
13/04/2017 NOVOZYMES PRESENTATION Rice starch Cotton CFT CS-28 Polyester CFT PS-28 Nylon CFT NS-28 Polyacryl CFT PAS-28 Cotton/Polyester CFT PCS-28 With Termamyl® With Stainzyme® Commercial powder with bleaching agent, dosage: 4 g/l, Commercial washing machine, temperature: 40°C, short programme

Status for Amylases Amylases important for stain removal in laundry
13/04/2017 NOVOZYMES PRESENTATION Status for Amylases Amylases important for stain removal in laundry Amylases are to a large extent recognised as a provider of basic detergency in laundry Amylases very important type of enzymes for dishwashing applications Stainzyme launch is a break through in amylase technology

Cellulases from Novozymes
13/04/2017 NOVOZYMES PRESENTATION Cellulases from Novozymes Celluzyme® first cellulase from NZ, colour clarification, prevents anti-redeposition, tolerant towards LAS. Multi-component enzyme product Carezyme® colour clarification, care cellulase, high activity on cellulose fibres, sensitive to LAS Endolase® prevents anti-redeposition and maintain whiteness (in EU liquids). Works on soluble cellulose Renozyme® colour clarification, robust towards anionic surfactants, cost effective cellulase

Many cellulases have a domain structure
13/04/2017 NOVOZYMES PRESENTATION Many cellulases have a domain structure Catalytic Core Hydrophilic Linker Cellulose Binding Domain

“Cellulase” enzymes working together to degrade cellulose
13/04/2017 NOVOZYMES PRESENTATION “Cellulase” enzymes working together to degrade cellulose CBH II Carezyme / Renozyme EG G R NR Cellulase: a mix of all activities – difficult to control Carezyme – color clarification / Care cellulase – it is the EGV activity from celluzyme – working on insoluble cellulose, CBD, therefor sensitive to proteases as they might cut of the CBD in liquid detergents Renoozyme: Carezyme like cellulase, has the CBD, cost effective variant of carezyme, developing markeds – Endolase: anti-redeposition cellulase, no colar car effects, only working on soluble cellulose, no CBD, good in liquids, good under EU conditions due to low water amount compared to eg US conditions. EG EG CBH I Endolase NOVOZYMES PRESENTATION NOVOZYMES A/S

13/04/2017 NOVOZYMES PRESENTATION Cellulase reactions Cellulases hydrolyse b-1,4-glucose-linkages forming soluble dextrins The substrate is water-rich amorphous cellulose In fibers this is especially found in stress points where the cellulose crystal structure is disrupted Some cellulases are only active on soluble forms of cellulose

Removal of fuzz = Clearer Colors With Carezyme No Cellulase 13/04/2017
NOVOZYMES PRESENTATION Removal of fuzz = Clearer Colors With Carezyme No Cellulase

Cellulase effects Fuzz removal (Color clarification)
13/04/2017 NOVOZYMES PRESENTATION Cellulase effects Fuzz removal (Color clarification) Depilling / anti-pilling Soil removal (cleaning) Anti-redeposition Softness

Endolase® antiredeposition effect
13/04/2017 NOVOZYMES PRESENTATION Endolase® antiredeposition effect Without cellulase White cotton “Grey” cotton + Wash With cellulase Carbon soil “Less grey ” cotton

Endolase® cleaning effects
13/04/2017 NOVOZYMES PRESENTATION Endolase® cleaning effects Whiteness No Endolase Endolase Unwashed Wash 1 Wash 2 Wash 3

13/04/2017 NOVOZYMES PRESENTATION Status for cellulases Big difference in effects of different cellulases Color clarification & care: Celluzyme, Carezyme and Renozyme Anti-redeposition & Cleaning: Celluzyme and Endolase Cellulases can support both basic performance and aesthetics Anti redeposition: Celluzyme delivers both care and anti-redeposition effect as it delivers the whole cellulase complex (multicomponent), however it is more difficult to control due to the EG5 activitiy (Carezyme). Renozyme was produced as a cost efficient cellulase – has same effect as Carezyme however it is more tolerant towards LAS. Developed for developing markets where the window for enzymes is smal. NOVOZYMES PRESENTATION NOVOZYMES A/S

Factors influencing enzyme wash performance
13/04/2017 NOVOZYMES PRESENTATION Factors influencing enzyme wash performance Detergent composition Enzyme concentration Temperature pH Water hardness Contact time Type of soil Type of wash system Type of wash method

Thank you! Slide Divider guide
13/04/2017 NOVOZYMES PRESENTATION Slide Divider Use: Drawing attention to the different chapters or themes in the presentation Edit: Change the photo by marking the background plane (largest object) in the grouped object: > Click on Format AutoShape > Under Fill – Colour, choose Fill Effects > Go to Picture tab and click on Select Picture tab > Choose picture > Click OK guide Thank you!

Enzymes used in the detergent industry

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