A Project on Application of cellulase enzyme on textile Processing

Submitted by: Name
Arup Chandra Roy Md. Iqbal Shaikh Mehedi Hasan Khan Abu Sayed Gm. Mortuza Md. Asadul Islam Md. Sharif Miah Sufal Chandra Sutradhar Md.Abdus Sobahan

072-178-041 072-223-041 072-224-041 072-226-041 073-050-741 081-028-741 081-118-041 081-272-041 081-279-041

Md. Mahbub Hasan Sr. Lecturer , department of textile Engineering Primeasia University

Adessertation submmited on: “Application of cellulase enzyme on textile Processing” To the department of Textile engineering in partial fulfillment of the credit requirement for achiving the Bachalor Degree in textile engineering by PRIMEASIA UNVERSITY. Submitted by: Name
Arup Chandra Roy Md. Iqbal Shaikh Mehedi Hasan Khan Abu Sayed Gm. Mortuza Md. Asadul Islam Md. Sharif Miah Sufal Chandra Sutradhar Md.Abdus Sobahan

072-178-041 072-223-041 072-224-041 072-226-041 073-050-741 081-028-741 081-118-041 081-272-041 081-279-041

Md. Mahbub Hasan Sr. Lecturer , department of textile Engineering Primeasia University

“Application of Cellulase enzyme on textile processing”


By the name of Allmighty ALLAH; The most gracious; The most merciful; The lord of all creation.


To our Parents


We would like to express our gratitude to all those who gave us the possibility to complete this project. We want to thank the department of textile engineering for giving the permission to commence the project at first instance, to do the necessary research work and to use departmental data. We have further more to thank the supervising teacher Md. Mahabub Hasan, Sr. lecturer of textile Engg. Dept. who gave and confirmed this permission and encourage us to go ahead with our project. We are deeply indebted to Rahamot Group of Industries Limited, Hams Fashions Limited, Jamuna Group of Industries Limited, Fakir Knitwear Limited, Graphics Textiles Limited, BASF Chemicals Limited and Hotapara Garments Limited for their help and co-operation and also to the Primeasia University faculties. Their help, stimulating, suggestions and encouragement helped us in all the time for this research and for writing the project. At last all the credits go to Almighty Allah, who enabled us to complete the project.


Now a day’s the whole universe is concern about the hygiene and they are looking for eco-friendly processes every where. Textile processing is one of the most pollutants responsible for polluting the environment at a large degree. Use of Cellulase enzyme for denim washing & Bio-polishing of knit fabric is a standard eco-friendly technique to achieve desired appearance and washing of denim and also the desired appearance of the knit fabric. Applications of enzymes to replace harsh chemicals and other difficulties for process industries have been practiced for decades. In this study we have tried to observe the washing effect of denim fabric and the bio-polishing effect of knit fabric with Cellulase enzyme under different conditions. This work also evaluates the effect of cellulase finishing of dyed cotton fabrics.


Objectives of the Project: The technology which is used to modify the appearance, out look, comfortability & fashion of the garments is called garment washing.

To know the effect of Cellulase enzyme on fabric or garments after fabric or garments washing.  To find out the washing technique by which faded/old look effect is created in the garments.  To create wash look appearance by cellulase washing of garments. After washing the garments create a new look which seems the new touch of fashion. To produce soft garments by enzyme washing then it seems to best touch of garments. To attract the customers/Buyer by different types of Fashionable washing and market developments. To produce shrinkage free garments by enzyme washing.  To remove dirt, dust, spot by enzyme washing and create new wash down appearance at the same time. To obtain the best Bio-polishing effect on the knit fabric.


Scope: Enzyme wash is suitable for all cotton/ denim garments, cellulosic woven or knitted and regenerated cellulosic like tencel and modal etc.  Permanent faded, soft hand effect can be produced on the treated material.  It facilitates the removal of dead / immature cotton and reduction of any fibre hairs on the surface (fewer tendencies to pilling).  Faded look or wash-down look can be produced on treated material.


  

Possibility of producing same effect is difficult. Possibility of fabric damages during washing. Application of cellulase enzyme in the textile processing often results in a certain amount of loss of tensile strength along with the desired performance.




Visited to different washing Industries and knit dyeing industries ↓ Observed Different processes of denim washing (cellulase enzyme) and Bio-polishing ↓ Observed the effect of enzyme on fabric and garments. ↓ Collected the Samples ↓ Group discussion done ↓ Selected the suitable Process and Recipe of Enzyme application on Textile Processing

SECONDARY STAGES: Informations Collected from:     Books, internet Visiting industries Magazine Seminar And Completed


Chapter -1 Garments washing Types of washing Chapter-2 Enzyme ---------------------------------------------------------18-19 20 21 22 23-24 25-31 ----------------------------------------------------------------------------------------------------------------16 17

Etymology and history ----------------------------------------------------Enzymes are specific and work in mild conditions -------------------Sources of enzymes -------------------------------------------------------------------------------------------------------

Different type of enzymes

Types of machines used in garments washing plant -------------------



Cellulase wash ----------------------------------------------------------------Cellulase enzyme processing -------------------------------------------Process sequences of enzyme wash (for dark shade) ----------------Enzyme wash process (for light shade) -------------------------------Stone enzyme wash -----------------------------------------------------Enzyme inactivation ----------------------------------------------------Advantage of enzyme washing -----------------------------------------

32-34 35-36 37-47 48-51 52-55 56 57

Chapter-4 Bio-polishing of knit fabric --------------------------------------------58-59 60-61 62 63 64-67 68

Mechanism of bio-finishing -------------------------------------------Bio-polishing of knit fabric with cellulase enzyme -----------------Favorable conditions of enzyme for bio-finishing ------------------Procedure of bio-polishing of knit fabric -----------------------------Advantages of using enzymes for bio-polishing are -----------------


Cellulase enzyme offers the following benefits ---------------------Requirement of enzyme wash ------------------------------------------

69 70

Conclusion ----------------------------------------------------------------




The technology which is used to modify the appearance, out look, comfortability & fashion of the garments is called garment washing. The changes occur during washing due to the following actions:  Mechanical/rubbing/abrasion between garments to garments;  Garments to machine; and  Chemical action.

Garments washing is done due to the following reasons:
 Washing process of garment is done to create wash look appearance. After washing the garments create a new look which seems the new touch of fashion.  By the washing technique, faded/old look, color or tinted effect is created in the garments which also seems the best touch of garments.  The main and important function of washing is to reduce size materials as a result the garment become size free and become soft hand feel.  When these soft garments are touched then it seems to best touch of garments.  To attract the customers/Buyer by different types of Fashionable washing and market developments.  Due to washing, shrinkage occurs in the garments. There is no possibility of further shrinkage of the wash garments.  Any dirt, spot or germ if added in the garments during manufacturing is also removed due to washing.


 Enzyme wash (Bio Polish wash & enzyme wash)  Stone Enzyme wash  Stone wash  Bleach wash  Sand Blasting  Permanent wrinkle  Normal wash/Garments wash  Pigment wash  Caustic wash




Enzymes are proteins that catalyze (i.e. accelerate) chemical reactions. Like all catalysts, enzymes work by lowering the activation energy for a reaction and thus dramatically accelerating the rate of the reaction. However, enzymes do differ from most other catalysts by being much more specific. Enzymes are known to catalyze about 4,000 biochemical reactions. Enzymes are proteins produced by living organisms. All organisms produce a wide range of enzymes to accomplish necessary biological tasks. Some enzymes can also be replicated in the lab, or engineered to perform in a particular way. One of the reasons that enzyme washing is so ecologically friendly is the natural origins of enzymes, which biodegrade, rather than lingering in the water supply. Enzyme washing products are also much more potent than other laundry products, requiring people to use far less, in terms of volume. Enzymes, like other proteins, consist of long chains of amino acids held together by peptide bonds. They are present in all living cells, where they perform a vital function by controlling the metabolic processes, whereby nutrients are converted into energy and new cells. Enzymes are biocatalyst, and by their mere presence, and without being consumed in the process, enzymes can speed up chemical processes that would otherwise run very slowly. After the reaction is complete, the enzyme is released again, ready to start another reaction. In principle, this could go on forever, but in practically most catalysts have a limited stability, and over a period of time they lose, their activity and are not usable again. Generally, most enzymes are used only once and discarded after; they have done their job. Different types of enzymes are suitable for different stains. In all cases, the enzyme washing process breaks the stain down into smaller molecules which can be removed with water or conventional soap. Amylases will remove starch based laundry stains, while proteases break down protein chains, making them suitable

for protein stains. Lipases work very well on grease and oil, and cellulases are excellent general cleaners. Enzyme washing also yields a softer, more supple garment. We have worked only with cellulose enzyme. We tried to know the effect of cellulose enzyme on garments washing. For delicate garments, enzyme washing can be an excellent way to get clothing fresh and clean. Enzymes also work at very low temperatures, making them suitable for cold wash only things ranging from silk to wet suits. Many natural detergent products mix enzymes into their formulas, to ensure that they are effective at all temperatures and on all stains.


Etymology and history:
In 1878 German physiologist Wilhelm Kühne (1837–1900) coined the term enzyme, which comes from Greek ενζυμον "in leaven", to describe this process. The word enzyme was used later to refer to nonliving substances such as pepsin, and the word ferment used to refer to chemical activity produced by living organisms. Enzymes are usually named according to the reaction they carry out. Typically the suffix -ase is added to the name of the substrate (e.g., lactase is the enzyme that cleaves lactose) or the type of reaction (e.g., DNA polymerase forms DNA polymers). Having shown that enzymes could function outside a living cell, the next step was to determine their biochemical nature. Many early workers noted that enzymatic activity was associated with proteins, but several scientists (such as Nobel laureate Richard Willstätter) argued that proteins were merely carriers for the true enzymes and that proteins per se were incapable of catalysis. However, in 1926, James B. Sumner showed that the enzyme urease was a pure protein and crystallized it; Sumner did likewise for the enzyme catalase in 1937 The conclusion that pure proteins can be enzymes was definitively proved by Northrop and Stanley, who worked on the digestive enzymes pepsin (1930), trypsin and chymotrypsin. These three scientists were awarded the 1946 Nobel Prize in Chemistry.


Enzymes are specific and work in mild conditions:
Enzymes can work at atmospheric pressure and in mild conditions with respect to temperature and acidity (pH). Most enzymes function optimally at a temperature of 30 -700C and at PH values, which are near the neutral point (PH 7). Now-a-days, special enzymes have been developed that work at higher temperatures for specific applications. Enzyme processes are potentially energy saving and save investing in special equipment resistant to heat, pressure or corrosion. Enzymes, due to their efficiency, specific action, the mild conditions in which they work and their high biodegradability, they are very well suited for a wide range of industrial applications.

Enzymes are part of a sustainable environment:
Industrial enzymes can be produced in an ecologically sound way where the waste sludge is recycled as fertilizer.

Enzymes and industrial applications:
Industrial enzymes are originated from microorganisms in the soil. Microorganisms are usually bacteria, fungi or yeast. One microorganism contains over 1,000 different enzymes. A long period of trial and error in the laboratory is needed to isolate the best microorganism for producing a particular type of enzyme. When the right microorganism has been found, it has to be modified so that it is capable of producing the desired enzyme at high yields.

Enzymes for Textile:
There are a lot of enzymes like amylases, cellulases, catalase, pectinase and protease for various textile wet-processing applications like desizing, biopolishing, denim finishing, bleach clean-up, bio-scouring and de-wooling.


Sources of Enzymes:
According to Encyclopedia of Science & Technology (1992), industrial enzymes are obtained from three major sources: Plant: Malt diastase, papain, bromelain and ficin. Animal: Pancreatic enzymes, pepsin, lipase, catalase, and rennin. Microbial: Amylases, proteases, pectinases, invertase and glucose oxidase.

Enzymes are the products of living cells and like proteins; no enzyme has as yet been successfully synthesized. Among many sources these are the most common: animal tissues, organs and fluids such as muscle, liver and saliva; animal products, such as milk and eggs; vegetable materials and a variety of seeds as reported in Encyclopedia Americana (1967). Pancreatic enzymes are prepared from slaughter house waste such as pancrease, clotted blood, liver etc.


Different type of Enzymes: Proteases:
Proteases are the most widely used enzymes in the detergent industry. Proteases hydrolyze proteins and break them down into more soluble polypeptides or free amino acids. As a result of the combined effect of surfactants and enzymes, stubborn stains can be removed from fibres.

Though enzymes can easily digest protein stains, oily and fatty stains have always been troublesome to remove. The trend towards lower washing temperatures has made the removal of grease spots an even bigger problem. This applies particularly to materials made up of a blend of cotton and polyester. The lipase is capable of removing fatty stains such as fats, butter, salad oil, sauces and the tough stains on collars and cuffs.

Amylases are used to remove residues of starch-based foods like potatoes, spaghetti, custards, gravies and chocolate. This type of enzyme can be used in laundry detergents as well as in dishwashing detergents.

The development of detergent enzymes has mainly focused on enzymes capable of removing stains. However, a cellulase enzyme has properties enabling it to modify the structure of cellulose fibre on cotton and cotton blends. When it is added to a detergent, it results into the following effects: Colour brightening-When garments made of cotton or cotton blends have been washed several times, they tend to get a 'fluffy' look and the colours become duller. This effect is due to the formation of microfibrils that become partly detached from the main fibres. The light falling on the garment is reflected back to a greater extent giving the impression that the colour is duller. These fibrils, however, can be degraded by the cellulase enzyme, restoring a smooth surface to the fibre and restoring the garment to its original colour.

Softening- The enzyme also has a significant softening effect on the fabric, probably due to the removal of the microfibrils. Soil removal- Some dirt particles are trapped in the network of microfibrils and are released when the microfibrils are removed by the cellulase enzyme.


The precise properties of cellulases vary depending on their origin. The majority of microbial cellulases studied have been shown to be acidic proteins with a significant carbohydrate content.


Stability and storage:
The activity of cellulase preparations has been found to be completely destroyed after 10-15 minutes at 80 °C. Solutions of cellulase at pH 5-7 are stable for 24 hours at 4 °C. These products should be stored at 4 °C, in a dry place in tightly closed containers. If stored in this manner, lyophilized preparation is stable for several months without significant loss of activity.

Types of Machines used in Garments Washing Plant:
 Sample washing Machine (Horizontal / Vertical Type)  Washing Machine (Side loading)  Washing Machine (Front loading)  Hydro extractor Machine  Dryer Machine (Steam)  Dryer Machine (Gas)  Chemical Mixture Machine  Industrial Oven (Gas/Electric/ Steam)  Boiler  Submersible Pump  Steam chamber for crinkle  E.T.P (Effluent Treatment Plant)  Generator  Sand Blasting Gun  Sand Blasting chamber  Spray gun and dummy  Compressor


Sample washing Machine (Horizontal):

Washing Machine (Side loading):


Washing Machine (Front loading):


Hydro extractor Machine:

Dryer Machine (Steam)


Dryer Machine (Gas):

Chemical Mixture Machine:







Cellulase enzymes are natural proteins which are used in denim garment processing to get stone wash look on to the denim garments without using stones or by reducing the use of pumice stone. Cellulase attacks primarily on the surface of the cellulose fibre, leaving the interior of the fibre as it is, by removing the indigo present in the surface layer of fibre. These enzymes are different from the alpha amylase enzymes used for starch removal in that they are selective only to the cellulose and will not degrade starch. Under certain conditions, their ability to react with cellulose (cotton) will result in surface fiber removal (weight loss). This will give the garments a washed appearance and soft hand.

There are different forms and types of cellulase enzymes used in Bangladesh. – One is Acid Enzyme (liquid) and another is Neutral Enzyme. – Neutral Enzyme is two types - a) Powder form, b) Liquid form i.e., SL Enzyme

Acid stable Enzyme:
– Acid enzyme is slightly brown in color. – PH range is 4.5 to 5.5 during washing. – Temperature – Time required : 40°c - 55°c. : 20 to 60 mins.

– Enzyme effects come within short time. – Staining/Bleeding occurs more on garments. – Production high


Neutral stable Enzyme:
– Found in powder form. – Enzyme is slightly white powder form. – PH range is 6 to 7 during washing. – Temperature – Time required : 40°c - 60°c. : 40 to 70 mins.

– Enzyme effects come slowly. – Less staining/bleeding on garments. – With pumice stone give good effect/abrasion on garments. – In dark shade enzyme effect is good.

– SL enzyme is liquid form. – PH range is 6 to 7. – Temperature – Time required : 40°c - 60°c. : 45 to 80 mins.

– Enzyme effects come slowly. – Less staining/bleeding on garments. – With pumice stone the effect is better. – For dark shade enzyme effect is good.

Alkaline stable(not used widely)


Denim fabric is constructed of indigo dyed cotton warp yarns and natural undyed cotton filling yarns. The warp yarns are dyed by a special process that deposits the dye on the surface of the yarn, facilitating easy removal of the dye for a wash down appearance. The consumer can achieve this washdown appearance over time as a garment is repeatedly worn and washed, or it can be achieved immediately by cellulase enzyme treatment. In recent years, supplier variations in the composition of cellulase have provided more effective cellulases, reduced back-staining (redeposited indigo dye on the backside) of denim, reduced strength loss, and an expanded range of optimum processing conditions. Other types of fabrics, including lighter weight fabrics, can be cellulase enzyme treated to achieve a cleaner, smoother surface by removing surface cotton fibers. This effect will remain evident even after multiple home launderings.


A number of factors impact cellulase activity during processing, and these factors can be classified as mechanical or chemical:

CHEMICAL Cellulase Type Cellulase Amount Process Time Process Temperature Process PH Auxiliary Chemicals Pre-treatments

Equipment Type Equipment Design Process Time Garments wt. Liquor Ratio Machine Speed Auxiliary Products

To optimize the application of cellulase to fabrics or garments, consideration must be given to how these individual factors will collectively retard or accelerate cellulase activity. Mechanical action varies from machine to machine, depending upon the type and design. Most fabrics are processed in jets, jigs, or high-speed air-tumbling units for surface modifications. Overflow jets provide low mechanical action, requiring use of the more aggressive, standard acid cellulase. Soft-flow jets deliver more mechanical action, so either the modified acid or standard acid cellulase could be used. With the higher mechanical action present in the high-pressure jets or air-tumbling units, any one of the cellulases could be used. Garments are enzyme processed in rotary drum or paddle machines, typically used for garment washing and dyeing. The mechanical action of the garments in these machines, including garment-to-garment abrasion, further enhances surface fiber removal, color washdown, and softness. Process times for cellulase treatment typically range from 0.5-2.0 hours. Delicate, lightweight fabrics/garments are processed for shorter time periods.

When carefully controlled, the desired surface modifications can be achieved within 30 minutes, without excessive strength loss. Heavier fabrics, such as denim, normally require longer processing times at specific cellulase concentrations to achieve the standard washdown of color reduction, seam abrasion, and softness. Back staining of indigo denim is usually resolved by switching from standard acid to modified acid or neutral cellulase. Other options to reduce back staining include the use of anti-redeposition agents, balancing enzyme amount/time factors, or reducing the load size. Temperature control throughout the cellulase enzyme cycle is important for both quality control and reproducibility. Due to the narrow temperature range (normally 45-600C) for optimum cellulase utilization, most operations choose to run the procedure at just below the maximum temperature of 600C. Further reduction in temperature may be employed when lightweight fabrics are treated or when the cycle time is extended. Sometimes, lowering the reaction temperature will broaden the effective PH range, an attractive strategy when ideal conditions are not achievable. As with temperature, PH level affects cellulase performance. For each cellulase type, an optimum PH exists where activity is maximized. Precise control is difficult due to the many inherent variables, but operating within a realistic range is crucial for quality control and reproducibility. A buffer system of phosphate, citrate, or acetate is recommended to maintain PH during the entire cycle due to the typical release of alkalinity from garments during processing. Most textile operations process knit and woven fabrics at a 10:1 ratio of cellulase solution to fabric, while garments are treated at lower ratios of 5:1. Often times, lower ratios require less cellulase usage, but care must be taken to achieve uniform results. Often overlooked auxiliary chemicals can have a dramatic impact on the activity of cellulase. Strongly ionic substances reduce cellulase activity. Generally, nonionic surfactants tend to boost enzyme activity. Dyestuffs and fabric finishes may have an impact on cellulase enzyme activity, and allowances should be made for their effects. Vat dyes generally do not inhibit cellulase activity, but pigments bound with film-forming acrylics will block cellulase access, therefore extended time may be needed for acceptable washdown. Direct dyes, particularly those after treated with fixing agents, and reactive dyes inhibit cellulase activity. Fabric

finishes, such as finishes for wrinkle resistance, reduce the effectiveness of cellulase by limiting accessibility to the internal fiber structure. However, a large number of wrinkle-resistant garments are being successfully post-washed with cellulase.

Process Sequences of Enzyme Wash (For Dark Shade):
The enzyme washing process of a batch of 60 kg denim men's long pant (Trouser) are described below:  First Step : DESIZING: – – – – – – – – – Weight (80 pcs) Add water @ M : L = 1 : 9 Machine Running Temperature Add Desizing agent @ 0.6 g/l Add Detergent @ 0.8 g/l Time Drop the liquor Wash 1 time by cold water : 60 kg : 540 Litre : 60°c. : 324 gm. : 432 gm. : 10- 15 mins.

 Second Step :
Enzyme Wash: – – – – Add water @ M : L = 1 :8 Temperature Add Acetic Acid @ 0.3 g/l Add Anti Back staining @ 0.6 g/l

: 450 Litre : 45°c. : 135 CC. : 270 gm.

– – – – – –

Add Enzyme @ 2.0 g/l : 900 gm. Time (Depend upon the shade) : 40- 80 mins. Drain the liquor. Increase temperature to 90°c and run 1 minute (for enzyme killing). Drain the bath. Rinse Twice, each 3 to 5 minutes

 Third Step : SOFTENING: – Add water @ M : L = 1 : 8 – Add Acetic Acid @ 0.6 g/l – Cationic Softener @ 1 g/l – Temperature – Time – Drain the bath – Then unload the garments on trolley.  Fourth Step : Hydro-extractor Machine: – After unloading garments from the washing machine then they are sent to hydro-extractor machine to remove excess water from the washed garments : 450 Litre. : 270 gm. : 450 gm. : Room. : 10 to 15 mins.

 Fifth Step:
Drying Machine: – Load 60 kg garments to gas dryer.

– Temperature set at 75°c to 85°c. – Run about 40 mins. – After then run 10 mins in cold dryer. – Sixth Step: – After Complete drying, the garments are sent to quality Control section for quality checking and good ones are ready for delivery.


Raw Denim

Enzyme Wash (Dark Shade)

Raw Denim

Enzyme washed


Raw Denim

Enzyme Washed


Raw Denim

Enzyme Washed denim (Dark shade)


Raw Denim

Enzyme washed Denim


Raw Denim

Enzyme Washed


Raw Denim

Washed denim (Medium Shade)


Medium shade Garments


 The enzyme washing process of a batch of 60 kgs denim men's long pant (Trouser) are described below:  First Step : Desizing : – – – – – – – – – Lot weight (80 pcs) : 60 kg denim long pant. Add water @ M : L = 1 : 9 : 540 Litre Machine Running. Temperature : 60°c. Add Desizing agent @ 0.6 g/l : 324 gm. Add Detergent @ 0.8 g/l : 432 gm. Time :15- 25 mins. Drop the liquor. Wash 1 time by cold water

 Second Step : Enzyme Wash – – – – – – – – – Add water @ L : R = 1 : 8 : 450 Litre Temperature : 45°c. Add Acetic Acid @ 0.6 g/l : 270 gm. Add Anti Back staining @ 0.6 g/l : 270 gm. Add Acid Enzyme @ 2.0 g/l : 900 gm. Time (Depend on shade) : 35- 60 mins. Increase temperature to 90°c and run 1 minute (for enzyme killing). Drain the bath. Rinse Twice, 5 minutes for each

 Third Step : Bleaching: – Add water @ M : L = 1 : 7 – Temperature – Add bleaching agent @ 20 g/l – Add sodium bi carbonate @ 4 g/l – Time (Depend on shade) – Drop the liquor. – Rinse Twice, 3 minutes for each : 420 Litre : 50°c to 60°c . : 8400 gm. : 1680 gm.

: 12 to 20 mts.

 Fourth Step : Neutralization -1 - Add water @ M : L = 1 : 10 – Temperature – Add sodium meta-bisulphite @ 2 g/l – Time – Drain the liquor. – Rinse Twice, 3 minutes for each : 600 Litre : 50°c. : 1200 gm.

: 10-15 mins


 Fifth Step : Second bleach: – Add water @ M : L = 1 : 8 – Temperature – Add hydrogen peroxide @ 4 g/l – Add caustic soda @ 1 g/l – Add detergent @ 1 g/l – Time – Drain the liquor. – Rinse Twice, 3 minutes for each and Neutralization is needed for Second time.  Sixth Step : Softening: – Add water @ M : L = 1 : 8 – Add Acetic Acid @ 0.6 g/l – Cationic Softener @ 1 g/l – Temperature – Time – Drain the bath. – Then unload the garments on trolley. – Seventh Step : Hydro-extractor Machine

: 480 Litre : 60°c. : 1920 gm. : 480 gm. : 480 gm. : 10 mins.

: 450 Litre. : 270 gm. : 450 gm. : Cold. : 10 to 15 mins.

After unloading garments from the washing machine then they are sent to hydroextractor machine to remove excess water from the washed garments.

 Eighth Step :
Drying Machine: – Load 60 kg garments to Steam dryer. – Temperature set at 60°c to 75°c. – Run about 40 mins. – Then run 10 mins in cold dryer.

 Ninth Step :
After dryer garments are sent to quality section for quality checking and then garments are ready for delivery

Light shade is produced by enzyme wash

Now in Bangladesh most of the Denim garment is required STONE ENZYME WASH effect. It is most popular wash for Buyer. A process of stone enzyme wash of 60 kg batch of Denim Long Pant as mentioned below :

Pumic stone used in stone wahing

 First Step : Pre-treatment: Desizing – Batch Weight – Add water @ M : L = 1 : 9 – Start the machine. – Temperature – Add desizing agent : – Add detergent/Antistain : – Time – Drop the liquor

: 60 kg : 540 litre.

: 60°c 0.6 g/l : 324 gm.

1 g/l : 540 gm. : 15 to 25 mins.

 Second Step : Hot wash: – Add water @ M : L = 1 : 9 – Temperature – Time : 540 litre. : 60°c. : 5 mins

 Third Step: Stone Enzyme Wash: – Add water @ M : L = 1 : 8 : 480 litre.

– Add pumice stone @ ½ volume of garments. – Add Enzyme @ 1.50 g/l – Add Acetic Acid @ 0.6 g/l : 720 gms. : 288 gms.

– Add Anti stain agent @ 0.8 g/l : 384 gms. – Temperature – Time (Depend on shade) : 40°c to 50°c.. : 60 to 70 mts.

– Then temperature raise to 90°c for 1 minute. – Drop the liquor. – Rinse Twice, each for 3 minutes. – Then pumice stone out from washing machine


 Fourth Step : Bleaching: – Add water @ M : L = 1 : 8 – Machine running. – Add bleaching powder (k.c.i) @ 10 g/l – Add soda ash @ 5 gm/l – Temperature – Time (Depend on shade) – Drop the liquor – Rinse twice, each for 3 minutes  Fifth Step : Neutralization: – Add water @ M : L = 1 : 9 : 540 litre. : 4800 gms : 480 litre

: 2400 gms : 60°c : 10-15 m

– Add sodium meta-bisulphite @ 3 g/l : 1620 gms – Temperature – Time – Drop the liquor – Rinse for 5 minutes. : 40°c : 10- 12 m


 Sixth Step : Softening: – Add water @ M : L = 1 : 8 – Add Acetic Acid @ 0.6 g/l – Cationic softener @ 1 gm/l – Time – Drop the liquor – Unload the garments on trolley. – Seventh Step : Hydro-extraction Hydro-extraction of the garments are done to remove excess water from the washed garments.  Eighth Step : Drying : – Load 40 kg garments – Set temperature at 75°c to 85°c. – Run for 35 to 40 mins. – Then run 10 minutes for cold dry. – Ninth Step : Delivery After inspection garments are ready for delivery. : 480 litre. : 288 gms. : 480 gms. : 5 mins.


Enzyme Inactivation:
To prevent any damage of the fabric after the finishing operation it is very essential that the reaction be terminated at the end of treatment by enzyme inactivation . If the enzyme is not inactivated entirely then at the end of the reaction fibres get damaged and even extreme cases total destruction of the material may result. The enzyme inactivation is therefore of great importance from the technical point of view. There are two distinct process of termination of enzyme: 1) Hot treatment at 800C for 20 minutes. 2) By raising the pH to 11-12.


Advantage of enzyme washing:
1. Soft handle and attractive clean appearance is obtained without severe damage to the surface of yarn. 2. Inexpensive, low-grade fabric quality can be finished to a top quality product by the removal of hairiness fluff and pills, etc. 3. Simple process handling and minimum effluent problem. 4. Better feel to touch and increased gloss or luster. 5. Prevents tendency of pilling after relatively short period of wear. 6. Can be applied on cellulose and its blend. 7. Due to mild condition of treatment process is less corrosive. 8. Fancy colour-flenced surface can be obtained without or a partial use of stone. 9. More reproducible effect can be obtained. 10.It allows more loading of the garment into machines. 11.Environmental friendly treatment. 12.Less damage to seam edges and badges. 13.Wear and tear of equipment is minimum due to absence of stone. 14.Use of softener can be avoided or minimised. 15.Easy handling of floor and severs as messy sludge of stones does not interfere. 16.Due to absence of stone, labour intensive operation of stone removal is not required. 17.Homogenous abrasion of the garments. 18.Puckering effect can also be obtained. 19.Problem of pumice powder contamination on garment is not there.

Evaluation of the Fabric:
Evaluation of the denim fabric after treatment with enzyme is, to be effective in reduction of hairiness, impartation of softness, wash-down effect etc. The enzyme was active in reducing hairiness, backstaining, wash-down effect, and imparting softness to the fabric or garments.


Bio-polishing of Knit Fabric:
Cotton and other natural and man-made cellulosic fibers develop pills or protruding fibers during spinning, weaving and wet processing operations, due to abrasions. These pills make the appearance of the yarn or fabric to be dull and hazy. In order to improve the appearance it is necessary to either remove the protruding fibers or to cement it to the surface properly by the application of some over coating. Cotton and other natural and man-made cellulosic fibers can be improved by an enzymatic treatment called Bio-Polishing. The main advantage of Bio-Polishing is the prevention of pilling. Enzyme cellulases hydrolyze the micro fibrils (hairs or fuzz or pills) protruding from the surface of yarn because they are most susceptible to enzymatic attack. This weakens the micro fibrils, which tend to break off from the main body of the fiber and leave a smoother yarn surface. After Bio-Polishing, the fabric shows a much lower pilling tendency. Other benefits of removing fuzz are a softer, smoother feel and superior brightness of the dyed shade or white. Unlike conventional softeners, which tend to be washed out and often result in a greasy feel, the softness-enhancing effects of Bio-Polishing are wash proof and non-greasy. For cotton fabrics, the use of Bio-Polishing is optional for upgrading the fabric. However, Bio-Polishing is almost essential for the new polynosic fiber lyocell. Lyocell is made from wood pulp and is characterized by a tendency to fibrillate easily when wet. In simple terms, fibrils on the surface of the fiber peel up. If they are not removed, finished garments made with lyocell will end up covered in pills. This is the reason why lyocell fabric is treated with cellulases during finishing. Cellulases also enhance the attractive, silky appearance of lyocell.


Bio-Polishing has a lasting effect on knitted as well as woven fabrics, giving improved resistance to pilling, a clearer, lint-free and fuzzless surface structure, and improved drapability and softness. Cellulase enzyme is used for bio-polishing of cellulosic fabric under acidic conditions. It partially digests excess and protruding yarns, loosening them from the fabric. The resulting fuzz is then easily removed by mechanical agitation of the fabric. This not only creates a smoother fabric with resistance to pilling, but also improves softness, luster and drape.


Mechanism of Biofinishing:
Understanding the behavior of the cellulolytic enzymes towards the cotton fiber is essential to minimize the related negative effect on the strength of the cotton fabric. Cellulose is made up of highly ordered crystalline region and less structured amorphous region. The degree of departure from crystallinity is variable to form the less structured purely amorphous region with all degrees of order in between. The amorphous region is generally responsible to form the fuzz and pilling in cotton fabric. Hence, a single type of cellulose preferentially endoglucanase may probably be sufficient for degradation of amorphous cellulose with minimum weight loss.

There is also evidence that during biosynthesis of cellulose the associated hemicelluloses regulate the aggregation process. Thus, application of cellulase for biofinishing along with xylanases may probably have an added advantage mainly in removal of the surface fibrils, small protruding fibers, seed coat fragments, and other natural impurities of cotton fiber. Use of xylanase along with cellulase in optimum concentration at suitable conditions for enzymatic action will prove to be beneficial with minimum harm to the fabric during biofinishing. We represent here a schematic diagram that indicates the probable mechanism of enzymatic finishing and stone washing of denim garment by endoglucanase Fig. During indigo staining of denim garment the indigo dye particles get adhered to the microfibrils that are present on the surface of the garment (Fig. b). Endoglucanase

acts on the amorphous region of the cellulose forming the protruding hairs on the fabric generally responsible for causing fuzz and pilling on the fabric as seen in the scanning electron micrograph and loosens it. The mechanical action in the washing machine in turn removes the loosened fibers to give a final finished product (Figs. d and b). Also as the protruding fibers are removed the indigo particles adhering to the surface of the protruding fibers are also removed thus giving the fabric a patchy appearance which is preferred (Fig. d). Xylanases will act on the seed coat fragments and other natural impurities thus giving a final finished touch to the cotton fabric. Cellulases are also used to improve the appearance of cellulosic fabrics by removing fuzz fiber and pills from the surface, reducing pilling propensity, or delivering softening benefits. Since cellulose is made up of highly ordered crystalline region and less structured amorphous region, a single type of cellulase preferentially endoglucanase will be sufficient for degradation of amorphous cellulose causing fuzz and piling of the fabric. Treatment of cotton fabric with cellulases along with an appropriate dose of xylanase will help in removal of seed coat fragments and other natural impurities of cotton.


Bio-Polishing of knit fabric with Cellulase Enzyme:
In the recent years enzymes have found a variety of uses in textile applications. Popular uses are stone washing of denims and surface modification of cellulosic fabrics to improve their appearance and handle. The process of treating with cellulases is termed as bio-polishing. In case of denims one can get stonewash effect without using pumice stones by using enzymes. Another advantage of using enzymes is that, these are environment friendly, since they are readily biodegradable. Besides, they will not leave chemical residue on the processed materials and the colour changes on the dyed goods are very less. Knitted goods treated with enzymes are free from surface hairiness and neps with much improved handle and flexibility. The fabric surface becomes smoother and more lustrous. There is also a lower tendency to further pilling possibly due to the fact that there are less protruding fibre ends from the yarns after the enzyme application. Bio-finishing also called bio-polishing, is a finishing process applied to cellulose textiles that produces permanent effects by the use of enzymes. Bio-finishing removes protruding fibres and slubs from fabrics, significantly reduces pilling, softens fabric hand and provides a smooth fabric appearance, especially for knitwear and as a pretreatment for printing.


Favorable conditions of enzyme for Bio-finishing:
More than with other chemical reactions, the enzyme catalysed hydrolysis of cellulose is strongly influenced by factors such as pH, temperature, time and agitation. The optimal pH for a particular cellulase depends upon its origin. Enzymes are more effective at pH 6 - 6.5. The reaction temperature is also critical since at low temperatures, the reaction rate is slower than desired, but very high temperature can deactivate the enzyme by providing enough energy to alter its molecular alignments and thereby destroy its catalystic ability. Since enzymes are true catalysts and are not consumed during the chemical reaction, the hydrolysis reaction will continue until either the reaction conditions change or the cellulose is physically removed from the reaction mixture.

Because the enzyme’s catalytic action is not reduced during the reaction, effective methods of ending the hydrolysis must be employed to prevent excessive fibre loss. Since the molecule’s physical alignments are crucial to its catalytic ability, procedures that alter the cellulase molecule’s internal structure can be used to deactivate the catalysis and halt the hydrolysis. High temperatures (> 700C or 1600F for at least 20 min or short drying at 1200C or 2480F), high PH (>10) and high electrolyte content as well as enzyme poisons can serve to terminate the reaction by distorting the enzyme’s molecular shape.

Recipe for Bio-finishing of knit fabric:
Neutral Cellulase Enzyme PH Time Temperature 55◦C 1% owf 5-6.5 1 hour


Procedure of bio-polishing of Knit Fabric:
• Load the fabric or garment in the machine • Fill the machine with water • Add nonionic wetting agents (0.2 to 0.3 gpl) • Adjust pH 4.5 to 5.5 with acetic acid • Add 2 gpl lubricant (non-ionic) • Run the machine for 30 minutes at 50-60 ◦C • Remove one garment from the machine and compare with the unwashed garment to see the effect of bio-polishing • If bio-polishing is satisfactory, raise the temperature gradually to 85◦C and maintain the temperature for 10 minutes to deactivate the enzyme • Drain the liquor • Cold rinse for 5-10 minutes followed by hydro extraction and tumble dry.


Process sequences of Bio-finishing:

Scouring & Bleaching


Enzyme process


After Treatment



Raw Fabric

Bio-polished Fabric

Recipe: Acid CellulasePHTimeTemperature0.8% owf 4-5.5 50-70 Minutes 55◦C

Raw sample

Bio-Polished sample (50 minutes)

Polished sample (70 minutes) Recipe: Acid CellulasePHTemperatureTime1% owf 4-5.5 55◦C 50-70 minutes.

Raw Fabric

Polished Fabric(1%)

Polished Fabric (1.5%) Recipe: Acid CellulasePHTemperatureTime1-1.5% owf 4-5.5 55◦C 50minutes.


Advantages of using enzymes for bio-polishing are:

• Hairiness, fluffs and pills are removed. • Material sticking (the burr effect) is prevented. • Improved handle. • Achievement of surface smoothness and a clear structural appearance. • Improved lustre. • Material texture relaxation. • Increased flexibility and therefore a soft handle even with over endproducts and mercerized fabric. • Improved sewability. • Fast to washing, low pilling tendency, no napping in use, or during care operation. • Lustrous, soft, top quality with a fine, high quality surface appearance. Provides surface polishing and softening.


Cellulase enzyme offers the following benefits:
     Specific action on protruding fibers only. Improves texture, softness and appearance of fabric. Wide variety of application processes. Eco-friendly & Biodegradable. No damage to strength of fabric.

Disadvantages of this finishing technique:
• Loss in weight • Loss in strength


Testing & Analysis:
The following tests can be carried out to assess the properties of bio-polished cotton knitted fabric. i) Wash fastness , ii) Abrasion resistance, iii) Weight loss, iv) Pilling.

Requirement of Enzyme Wash:
Now we can say Enzyme wash is required for the following reasons: – To remove the size materials from the garments. – To remove the starch presents on the garments fabrics. – To achieve the high low abrasion (stone effect) on garment and seam abrasion in sewing area. – Enzyme attack as chemically not mechanically for this reason low damage/wastage than stone wash. – For soft feeling to wear the garment. – To achieve the buyer required sample. – To increase the color fastness & rubbing fastness. – Especially develop the "Bio-Polishing" effect of cotton/denim. – Enzyme improves the anti-pilling properties. – Enzyme attacks more the surface of the fabrics and gives a very smooth surface


The most attractive feature of denim fabric is its washing effect. This is produced by mechanical and chemical processing. The entire processes gives rise to the wastage hence effluent and environment is polluting at a large scale. Enzyme technology provides an environment friendly process to obtain the desired effect. The bio-polishing of knit fabric can also be done with cellulase enzyme which removes the protruding fibres of the fabric and gives a smoother, softer, Brighter and cleaner fabric. The activity of the Enzyme can be controlled easily by maintaining the PH, Temperature, Concentration and time. The enzyme washing and Bio-polishing provides a better and uniform effect over the conventional processes. Though the enzymes are costly but it is energy and time saving over the conventional processes. The limitation of Enzyme washing is that when localized washing effect is needed it is difficult to produce it. In the future, enzyme technology will continue in importance as biochemists and textile chemists strive to create new product and processing innovations to further revolutionize textile processing.



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