Enzymes used in the detergent industry 08/04/2015 2 08/04/2015 NOVOZYMES PRESENTATION Carbohydrases 4 Types of enzymes are used for different detergents Substrate Examples Protease Proteins Blood, egg, milk grass, meat Lipase Triglycerides Organic fats and oils, sebum, lipstick Amylase Starch Rice, noodles, potatoes, corn Cellulase Cellulose Cotton, viscose 3 08/04/2015 NOVOZYMES PRESENTATION Catalytic actions of detergent enzymes 4 08/04/2015 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 5 08/04/2015 NOVOZYMES PRESENTATION Removal of stain 1: The stain is bound to a surface Stain Substrate Fibre 6 08/04/2015 NOVOZYMES PRESENTATION Removal of stain 2: A surfactant has been added Surfactant Soil Fibre 7 08/04/2015 NOVOZYMES PRESENTATION Removal of stain 3: Enzymes are added and degrade available sites Enzyme Surfactant Soil Fibre 8 08/04/2015 NOVOZYMES PRESENTATION Removal of Stain 4: The soil comes loose and goes into suspension Surfactant Enzyme Soil Fibre 9 08/04/2015 NOVOZYMES PRESENTATION General principle of stain removal Clean vest Stain Wash, no enzýmes Stain less visible Wash with enzymes Stain no longer visible 10 08/04/2015 NOVOZYMES PRESENTATION Removal of invisible stains: redeposition Clean vest Stain Washed Washed in dirty water, No enzymes Washed in dirty water, With enzymes Conclusion: Enzymes give more permanent cleaning! 11 08/04/2015 NOVOZYMES PRESENTATION Removal of inorganic particulate soils H1: washed without cellulase H2: washed with cleaning cellulase 12 08/04/2015 NOVOZYMES PRESENTATION Removal of fuzz and pills Without cellulase With the care cellulase: Carezyme® 13 08/04/2015 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? 14 08/04/2015 NOVOZYMES PRESENTATION International brand support model Cellulase Lipase Amylase Protease Basic performance 15 08/04/2015 NOVOZYMES PRESENTATION International brand support model Polarzyme Stainzyme 16 08/04/2015 Proteases NOVOZYMES PRESENTATION 17 08/04/2015 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 18 08/04/2015 NOVOZYMES PRESENTATION Proteases catalyze the hydrolytic cleavage of the peptide chain Protease -R1CH-CO-NH-CR2H- + H2O → Proteases can provide: • stain removal • cleanliness • general whitening -CR1H-COO- + +H N-CR H3 2 19 08/04/2015 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 20 08/04/2015 NOVOZYMES PRESENTATION Protease effect on blood spots not washed washed 3 times, no protease washed 3 times, with protease 21 08/04/2015 NOVOZYMES PRESENTATION Stain removal with protease in laundry Protease no yes Protease no yes Cocoa (CFT) Grass (Equest) Blood (CFT) Spinach (Equest) Salmon paste (Equest) Olives (Equest) 22 08/04/2015 NOVOZYMES PRESENTATION Protease effect on blood spots Protease effect after 3 wash cycles and re-soiling No protease Re-soiling With Protease Re-soiling 23 08/04/2015 NOVOZYMES PRESENTATION Proteases perform well at high pH The figure shows Alcalase versus Savinase Relative Activity (%) pH 24 08/04/2015 NOVOZYMES PRESENTATION Wash performance of polarzyme 12T 25 °C, TOM1, Regular powder, 10 °dH, 20 min, EMPA 117 swatch Unwashed 1) TOM (Tergotometer) is a small scale method simulating top loading US washing machine 25 08/04/2015 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-8 Grass on cotton PCS-8 Grass on cotton/polyester EMPA164 Grass on cotton WfK 10N Egg on cotton WfK 20N Egg on cotton/polyester 26 08/04/2015 NOVOZYMES PRESENTATION Low temperature profile of polarzyme Activity profile of Polarzyme shifted towards lower temperatures maintaining activity at high temperature Activity temperature profile at pH 9 Relative activity 1,0 Polarzyme Savinase 0,8 0,6 0,4 0,2 0,0 0 20 40 60 Te mpe rature (°C) 80 100 27 08/04/2015 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 28 08/04/2015 NOVOZYMES PRESENTATION Protease benefit in wash & wear test - overall consumer preference 29 08/04/2015 NOVOZYMES PRESENTATION Protease benefit in wash & wear test consumer preference for each performance attribute 30 08/04/2015 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 31 08/04/2015 Lipases NOVOZYMES PRESENTATION 32 08/04/2015 NOVOZYMES PRESENTATION Lipase mechanism Lipase Lipase + No odor + Odor Short chain fatty acids (C4 –C8) Dairy soils 33 08/04/2015 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. 34 08/04/2015 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 - Lipase Give in-depth cleaning Reduces redeposition + Lipase General whitening Lipstick 35 08/04/2015 NOVOZYMES PRESENTATION Activity of Lipolase during drying Relative Activity (%) 36 08/04/2015 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 37 08/04/2015 NOVOZYMES PRESENTATION Lipase for fat removal from outside of cotton fibres no lipase Lard Sebum Olive Oil with lipase 38 08/04/2015 NOVOZYMES PRESENTATION Lipase effect on inside of cotton fibres Cross sectional specimen of yarns soiled with olive oil Olive oil Unwashed Olive oil Washed with detergent Olive oil Washed with detergent & lipase 39 08/04/2015 NOVOZYMES PRESENTATION Lipase 1’st wash effect on polyester 0.2 mm washed without lipase 0.2 mm washed with lipase 40 08/04/2015 NOVOZYMES PRESENTATION Lipex - visual soil removal in 1st wash No Lipex Lipex 500 LU/L Lipex 1000LU/L 41 08/04/2015 NOVOZYMES PRESENTATION Anti-redeposition benefits after 1st wash - a secondary effect of Lipases Unwashed textiles - Lipex + Lipex 42 08/04/2015 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 43 08/04/2015 Amylases NOVOZYMES PRESENTATION 44 08/04/2015 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 45 08/04/2015 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 46 08/04/2015 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 47 08/04/2015 NOVOZYMES PRESENTATION Removal of invisible starch stain Clean vest Starch stain Washed in dirty water, No enzymes Washed in any detergent Washed in dirty water, With enzymes 48 08/04/2015 NOVOZYMES PRESENTATION pH-profile of Termamyl and Stainzyme 49 08/04/2015 NOVOZYMES PRESENTATION Temp. profile for Termamyl and Stainzyme 50 08/04/2015 NOVOZYMES PRESENTATION Washing time 51 08/04/2015 NOVOZYMES PRESENTATION Different pH profiles 52 08/04/2015 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 53 08/04/2015 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 54 08/04/2015 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 55 08/04/2015 NOVOZYMES PRESENTATION Cellulases from Novozymes Celluzyme® first cellulase from NZ, colour clarification, prevents anti-redeposition, tolerant towards LAS. Multi-component enzyme product. 1986. Carezyme® colour clarification, care cellulase, high activity on cellulose fibres, sensitive to LAS. 1998. Endolase® prevents anti-redeposition and maintain whiteness (in EU liquids). Works on soluble cellulose. 1999. Renozyme® colour clarification, robust towards anionic surfactants, cost effective cellulase 56 08/04/2015 NOVOZYMES PRESENTATION Many cellulases have a domain structure Catalytic Core Hydrophilic Linker Cellulose Binding Domain 57 08/04/2015 NOVOZYMES PRESENTATION “Cellulase” enzymes working together to degrade cellulose Carezyme / Renozyme CBH II EG G R NR EG Endolase EG CBH I 58 08/04/2015 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 59 08/04/2015 NOVOZYMES PRESENTATION Removal of fuzz = Clearer Colors No Cellulase With Carezyme 60 08/04/2015 NOVOZYMES PRESENTATION Cellulase effects Fuzz removal (Color clarification) Depilling / anti-pilling Soil removal (cleaning) Anti-redeposition Softness 61 08/04/2015 NOVOZYMES PRESENTATION Endolase® antiredeposition effect Without cellulase White cotton “Grey” cotton + Wash With cellulase Carbon soil “Less grey ” cotton 62 08/04/2015 NOVOZYMES PRESENTATION Endolase® cleaning effects No Endolase Whiteness Unwashed Wash 1 Wash 2 Endolase Wash 3 63 08/04/2015 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 64 08/04/2015 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 65 08/04/2015 NOVOZYMES PRESENTATION Thank you!