ECO-FRIENDLY PRETREATMENTS

INTRODUCTION

The fabric should be free from natural & added impurities before it goes for coloration i.e.
dyeing or printing. Some of the chemicals like caustic soda, soda ash, hydrogen peroxide,
hydrochloric acid, detergents and auxiliaries that are used at different stages preparatory process
to remove such an impurities are found to be harmful to the environment. Caustic soda and soda
ash though are safe chemicals; but they cause high TDS in effluent. Detergent and surfactant add
Phosphorus to the outlet and require special treatment. They are also not easily biodegradable.
Auxiliaries like wetting agent contain phenol, which are not removed easily in the course of
effluent treatment. Modern wet processing industries are using the enzymes in the preparatory
process instead of using harmful chemical because enzyme are more convenient, effective and
environment friendly.

ENZYME APPLICATIONS IN TEXTILE PREPARATORY PROCESS

Enzymatic desizing:-
Using amylase enzymes for the removal of starch sizes is one of the oldest enzyme applications.
Amylases are enzymes which hydrolyse starch molecules to give diverse products, including
dextrins and progressively smaller polymers composed of glucose units. (it is carried out at the
temperature of 30-60°C, optimum pH is 5.5-6.5.) These partly degraded oligosaccarides cannot
be reused and are usually discharged, contributing large amounts of Chemical Oxygen Demand
(COD) and Biological Chemical Oxygen Demand (BOD) to effluent streams. 50-80% of the
COD in the effluents of textile finishing industries is caused by sizing agents.

 Enzymatic desizing and Glucose Generation in Desizing Liquor-

There are basically four groups of starch-converting enzymes: (i) endoamylases; (ii)
exoamylases; (iii) debranching enzymes; and (iv) transferases. Endoamylases are able to cleave
α,1-4 glycosidic bonds present in the inner part of the amylose or amylopectin chain. α-Amylase
is a well-known endoamylase. The end products of α-amylase action are oligosaccharides.
Enzymes belonging to the second group, the exoamylases, either exclusively cleave α,1-4
glycosidic bonds such as β-amylase, or cleave both α,1-4 and α,1-6 glycosidic bonds like
amyloglucosidase or glucoamylase. Exoamylases act on the external glucose residues of amylose
or amylopectin and thus produce only glucose (glucoamylase and α-glucosidase), maltose or
dextrin (β-amylase). The third group of starch-converting enzymes are the debranching ones that
exclusively hydrolyse α,1-6 glycosidic bonds: isoamylase and pullanase. The fourth group of
starch-converting enzymes are transferases, which cleave the α,1-4 glycosidic bond of the donor
molecule and transfer part of the donor to a glycosidic acceptor with the formation of a new
glycosidic bond.

An enzymatic process is proposed to utilise desizing baths for bleaching in which glucose
oxidase (GOx) enzymes generate hydrogen peroxide and gluconic acid using glucose as a
substrate. Advantages of the process are reducing the COD of the effluents by degrading glucose
units, and reducing the use of peroxide stabilising agents with the help of gluconic acid, which is
capable of complexing catalysts as well as saving water and energy by using desizing liquor for
bleach.

Conventional commercial desizing enzymes do not seem appropriate for this purpose since most
include α-amylase in formulations, whereas amyloglucosidases are suitable amylase enzymes to
degrade starch until it becomes glucose.
The performance of a commercial enzyme (an amyloglucosidase/ pullanase mixture) was tested
and process optimisation trials were performed. ( Aniş P., Davulcu A., Eren H. A.; Enzymatic
Pre-Treatment of Cotton. Part 1. Desizing and Glucose Generation in Desizing Liquor. FIBRES
& TEXTILES in Eastern Europe 2008, Vol. 16, No. 4 (69) pp. 100-103.) Optimum
circumstances obtained were: 0.75% (o.w.f.) enzyme, pH 4.1, 62 °C and a process time of 45
minutes.

Enzymatic scouring:
The objective of a scouring process is to make the material hydrophilic, before it undergoes other
processes like bleaching, dyeing and printing.

The conventional scouring operation consists of treating the cotton goods with 1- 2% of NaOH
solution at high pressure (lSlbs/in2) and temperature (121°C) for 4-5 hours. The above treatment
is not only energy intensive but also leads to environmental pollution. It is estimated that
scouring operation alone consumes about 1% of the total water used, contributes about 54% to
the total BOD and is responsible for 10-25% of the total pollution load generated during the
entire textile processing operation

Different enzymes like pectinases such as lyases, polygalacturonase endo acting type and
polygalacturonase exo acting type, proteases, cellulases such as endoglucanases,
cellobiohydrolases, xylanases, lipases and recently cutinases have been examined to degrade and
subsequently remove the natural component present in the outer layer of cotton fibres.

Lipases, were found to be less effective in fulfilling this task. Proteases were found to be
efficient to improve whiteness rather than hydrophilicity. Cellulases were the only enzymes
reported to improve the wet ability efficiently when applied without any other treatment or in
combination with other enzymes. However, cellulase also cause a decrease in fibre strength and
hence a decrease in fabric quality. The best results have been obtained by alkaline pectinases or
pectinases in combination with cellulase.

A typical adopted approach in past towards enzymatic scouring process of cotton

Fabrics

Still this process faces several problems,

 A need for long incubation time
 High enzyme doses
 Sometimes non-uniform enzyme action
 Uneven dyeing behavior.
 High temperature water treatment before enzyme incubation and
 Overall slow process speeds.

COMPARISON BETWEEN BIOSCOURING& ALKALINE SCOURING:

Test Parameter Bio Scouring Alkaline Scouring

PH 8 – 9.5 13-14

Temperature 50-600C/ extract 75-950C [95-1000C] 1300C

Residual Pectin 22-30% 10-12%

Weight loss <1.5% 3-8%

Handle very soft Harsh, Papery

Rinse Water 25-50% 100%

TDS [Total dissolved 25-50% 100%

solids]
 BOD 20 - 40% 100%

COD 20 - 45% 100%

 Bioscouring with pectinases-

It is found by many researchers that pectinase treatment alone results in adequate wettability.
Others have found out only little improvement in water absorbency. Pectinase enzymes are of 2
types- acidic and alkaline.
Both are found to be equally efficient in terms of improved absorbency as the conventional one.
The alkaline pectinase has similar performance to the acidic; however the former works on lower
concentration, which is beneficial from the economic point of view. However there is a better
absorbency and whiteness after treatment with alkaline pectinase than the acidic one.
The pectin content of cotton can be decreased by about 30% during the pectinase treatment. The
enzymatic treatment has no effect on the tensile strength, moreover it enhances the whiteness.

Whiteness of the bleached fabric-

 Bioscouring with cellulases-

This improves significantly the water wettability and water retention capacity of cotton fabrics.
Comparing the cellulase enzyme with the non-cellulosic ones, it is obvious that that the cellulase
causes greater weight loss and decreases the yarn tenacity. Morphology of the fiber changes
significantly. Cracks are more frequently observed in cellulase treated fabric than the non-
cellolosic treated one.

 Bioscouring with protease-

Protease are the enzyme that catalyse the hydrolysis of proteins. Results of many researches
prove that they have only a slight effect on improving wettability, however 50% of protein can
be removed from the fiber.
Protease appears to be effective in removing the proteinaceous materials only from the lumen but
not from the surface. Proteins of the surface layers are imbedded beneath the pectin material so
they are inaccessible for enzymes. However some proteases have been found out which also
removes the surface proteins.
There is no change in tensile strength and surface friction. However some proteases improved
lightness of the fabric.

 Biocsouring with lipases-

Lipase catalyse the hydrolysis of fats. Common fats are the esters of glycerol and fatty acids.
Lipase attack the ester bonds regenerating the water soluble glycerol and water insoluble fatty
acid. Lipase treatment does not improve water wetting and retention property of cotton. It may
be explained by either the low accessibility of waxes in the fibers, or the less susceptibility of
waxes to the enzymatic action. Lipase treatment results in reduced friction coefficient but does
not affect the yarn strength.

 Use of cutinase and pectinase in bio scouring –

Sufficient hydrophilicity during cotton scouring could be achieved by removing the outermost
waxy layer, followed by destabilizing the primary wall. Therefore, wax and pectin removal are
considered the most important steps in the enzymatic scouring process.

Cutinase: an enzyme for cotton wax removal

There is need to have an environmental friendly process that should degrade waxes at low
temperature and in an aqueous environment. An enzyme that can hydrolyse the ester bonds of
triglycerides and cutin, to destabilize the waxy layer in cotton fibre is desired. Enzymes from the
carboxylic ester hydrolyze group are promising candidates. Esterase, lipase and cutinase belong
to this group.

Lipases and cutinase are all lipolytic enzymes that show a high activity towards the substrate in
their aggregated form, hence they are suitable candidates for the cotton wax degradation.
Cutinase has some advantages over lipases for cotton wax degradation.
 Lipases, in general, require interfacial activation at the lipid water interface, whereas
cutinase does not. The interfacial activation is an extra energy need to expose the active
site of Lipase enzymes to the substrate.
 Another important characteristic of cutinase is that, it can hydrolyze waxes in the absence
of Ca2+ ions. Lipases need Ca2+ ions for their hydrolytic action. Since it is known that
the presence of Ca2+ ions can interfere negatively with the pectinase performance, it will
be difficult to combine lipases with pectinase during enzymatic cotton scouring.

Pectinases for primary wall destabilization-

Pectinase can degrade pectins in cotton fibres. Pectinolytic enzymes are produced by many plant-
associated micro organisms.
Polygalacturonases (acidic pectinase) and pectate lyase (alkaline pectinases) are promising
candidates for cotton scouring.
Bioprep 3000L (Novozymes) can be used as an alkaline pectinases.

Scouring parameters-

Pectinase concentration
The pectin removal increases with increasing pectinase concentration The maximum pectinase
concentration to be taken is 13 U/g ( depending upon the pectin content in the fabric), carried at
50°C for 5 minutes in a 50 mM Tris-HCl buffer at pH 8 with mild shaking. The purpose of the
mild shaking was to distribute the pectinase uniformly in the bulk medium and not to create
mechanical shear in the system.

pH
45% of the pectin can be removed when the pH was between 8 and 9. The pectin removal
efficiency of pectinase reduced drastically above pH 10 (20% pectin removal)

Temperature
According to Novozymes the optimum operating temperature range for the Bioprep 3000L was
50°C to 60°C

Ionic strength
The ionic strength of the Tris HCL buffer solution (pH 8) was selected as 50 mM. The pectin
removal increases with increasing ionic strength of the buffer solution.

Chelators
The use of chelators was desired in cotton scouring for the removal of Ca2+ ions from the
winding layer of cotton fibre. That will help to remove other non-cellulosic materials from the
primary wall and therefore a hydrophilic fabric. The efficient use of chelator at rinsing stage of
the scouring process has been justified in terms of maximum pectin removal. The chelator at this
stage can help to remove extra 10-15% of the pectin, which would not possible to remove
otherwise with the help of enzymes.

The general experimental setup for scouring experiment

Scouring experiments where performed in 1 L beaker in which three fabric samples of 5 × 12 cm
were treated in an enzyme solution of about 500 mL, with the 50 mM Tris-HCl buffer (pH 8).
The beaker was placed in a temperature controlled water bath at 30°C or 50°C. After the
treatment, the fabric samples were rinsed in 500 mL of water at 90°C for 15 minutes, to
inactivate the enzymes. Thereafter the samples were rinsed twice for 5 minutes in water at room
temperature. Finally, the samples were kept on a flat acrylic surface to be dried at air for at least
24 hours before evaluating the fabric samples.

Biobleaching :

Both conventional and enzymatic scouring processes do not affect natural colorants in the cotton
fiber to a major extent, and bleaching is essential for a good level of whiteness, especially for
material to be dyed in lighter shades. Currently, the most common industrial bleaching agent is
hydrogen peroxide, which is applied at pH 10.5-11 and temperatures close to the boil. Hydrogen
peroxide itself decomposes into environmentally benign compounds (water and oxygen),
however the conditions during bleaching pose a serious problem due to possible radical reactions
of the bleaching compounds with the fiber. These reactions can lead to a decrease in the degree
of polymerization and eventually to a drop in tensile strength, especially in presence of particular
metal ions which act as activators for hydrogen peroxide. Additionally, alkaline conditions
negatively influence the effluent treatment, and the temperatures needed for the process impact
the cost.

Different eco friendly approaches for enzymatic bleaching have been explored.

 Laccase/mediator systems –

Lactase (benzenediol:oxygen oxidoreductase; EC 1.10.3.2) is a blue multi-copper containing

oxidase and found in fungi growing on fruit, vegetables or trees, lignolytic white-rot fungus, and
bacteria.
Laccase/mediator systems have been used successfully for bleaching of wood pulp and it could
be expected that they are applicable to bleaching of cotton as well, although colorant
compositions in wood and cotton differ.

In the paper (L. Pereira, C. Bastos, T. Tzanov, A. Cavaco-Paulo, G.M Guebitz

‘Environmentally friendly bleaching of cotton using laccases’ Received: 5 May 2005 / Accepted:
9 August 2005 / Published online: 22 October 2005, it is seen for the first time that a laccase
from a newly isolated strain of T. hirsuta can oxidise flavonoids from cotton resulting in an
increase of whiteness.
Mediators such as ABTS (2-2-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) and HBT (1-
hydroxybenzo-triazole) are used.
Bleaching of cotton Cotton fabrics were first washed with a solution of Lutensol ON 30 in
deionised water (enough volume to cover the fibre) for 15 min at boiling temperature followed
by washing with tap water to remove the soap. Dried samples of 1 g were pre-treated with
laccase in 0.1 M, pH 5.0, acetate buffer at different conditions such as enzyme concentration,
bath volume, temperature, process duration, agitation level and addition of mediators. The blank
and control were done without enzyme and heat-inactivated laccase, respectively, at the same
conditions. After the pretreatment, cotton was washed with Lutensol as described above.
Bleaching was carried out for 1 h at 90◦C, with a solution containing (o.w.f.): 3.5% of silicate
(Na2/SiO2), 1% Na2CO3, 1% NaOH and 35% of 4% H2O2. Cotton was washed and dried.

Replacement of hydrogen peroxide for bleaching by an enzymatic bleaching system would not
only lead to better product quality due to less fibre damage but also to substantial savings of
washing water needed for removal of peroxide.

The major drawback of this bleaching system though is that suitable mediator compounds have
to be identified. Currently available mediators raise questions about efficiency and toxicity.
Mediators seem necessary for electron transfer to support the action of the fairly non-specific
laccases. They are consumed during the reaction and as such not considered true catalysts.

 Peroxidases-
This class of enzymes is generally capable of activating various oxidizing agents, one of which is
hydrogen peroxide. A fully satisfactory bleaching effect, however, has not yet been
accomplished with these enzymes. It is possible that their deactivation occurs too rapidly during
the bleaching process.

 Glucose oxidases-
These are enzymes, like peroxidases; belong to the class of oxidoreductases. Oxidoreductases are
a class of enzymes that oxidize or reduce a substrate by transfer of hydrogen(s) and/or
electron(s). Many of these enzymes are commonly known as oxidases, reductases, peroxidases,
hydrogenases, oxygenases, or dehydrogenases. Glucose oxidases generate hydrogen peroxide
and gluconic acid from glucose and oxygen. The use of this type of enzyme for bleaching would
allow for the reuse of sugar-contaminated effluent from other wet processing steps.
The Glucose Oxidase enzyme has to be added in the scoured (Amyloglucosidase) bath. In the
presence of molecular oxygen, glucose is oxidised by the enzyme glucose oxidase to gluconic
acid and hydrogen peroxide:
D- Gluconic acid is act as a sequestering agent during bleaching. To produce enough amount of
hydrogen peroxide in the bleaching bath more amount of Amyloglucosidase has to be added in
the scouring bath, there by enough glucose can be produced to obtain at least 250 mg per litter.

Whiteness of fabrics, bleached with enzymatically produced hydrogen peroxide, using

desizing baths as an additional source for glucose

 Bleaching of jute, cotton and jute/cotton blend using combined H 2O2 and potassium
per oxo-disulphate at room temp –

Bleaching of jute, cotton and its blend requires a high temp. As H 2O2 requires a high temperature
for its activity. Moreover the high temp causes oxidative degradation of the fiber. It is also
energy consuming.
When the fabric is treated with combined H2O2 and K2S2O8 , the later one acts as booster for the
former one and thus bleaching can be carried out at room temp ( 300C)
When the fabric is sequentially treated with 1% (owf) K 2S2O8 1 h, 300C followed by treatment
with 3% ( owf) H2O2, 1h, 300C, it shows a damage to jute fabric, loss in fabric tensile strength
and less improvement in the whiteness index.
However, when both the above steps were carried out simultaneously in a single bath, for 2 h at
300C with higher amount of alkali (2.5 %), a synergetic effect was seen with higher whiteness
index and less strength loss in jute.

Whiteness of fabrics, bleached with enzymatically produced hydrogen peroxide, using desizing
baths as an additional source for glucose,

enzymatic Enzymatic scouring + bleaching with

scouring reduced conc of H2O2 and alkali
Reduction in rinsing water 20 % 50 %
consumption

Reduction in BOD-load 20 % 40 %

Reduction in COD-load 20 % 40%

Combined scouring and bleaching of cotton using potassium per sulphate:

Hydrogen peroxide is used in the low temperature combined scouring and bleaching on the mill
grey mercerized cotton fabric at a lower temp in the presence of a bleach activator (potassium
persulphate) and an eco friendly stabilizer.
Potassium per sulphate (PPS) is known to activate the decomposition of H2O2. Complete
peroxide decomposition must be warranted to effectively bleach the cotton fabric and hence use
of stabilizer becomes necessary.
Here fabric is treated with alkaline H2O2 bath containing simultaneously PPS (potassium per
sulphate) as an activator and Contavan GAL of CHT (I) Ltd. as a commercial stabilizer based on
hydroxyl carbonic acid which is termed to be eco-friendly.
The low temp combined scouring and bleaching was carried out on grey mercerized fabric in
‘Mathis Labomat’ keeping MLR 1:30. After treatment the fabric is rinsed with fresh water and
dried.
From Taguchi analysis it is observed that change in temperature causes maximum variation in all
properties, whereas the change in concentration of H2O2 shows the least.

Temperature > Stabilizer GAL > Activator PPS > time > alkali NaOH > H2O2
From Taguchi analysis it can be seen that H2O2 shows the lowest effect on the mean result.
The highest effect of temp variations on the properties of the bleached fabric may be attributed to
the fact that the decomposition of PPS to generate free radicals, which in turn gives rise to
formation of peroxyl radicals is mainly governed by temperature.

Closed loop system (combined desizing scouring and bleaching):

In this, the main aim is to accomplish all aspects of preparatory finishing (desizing, scouring,
bleaching) by means of “green chemistry”, or in other words, non-toxic enzymatic treatments, in
a closed-loop procedure. The process is to be designed to allow for the treatment bath of
enzymatic desizing and bio scouring to be used for the bio bleaching step. In order to combine
treatments, optimum temperature and pH ranges of the involved enzymes had to be compatible.

Choice of Enzyme
For desizing, enzymes were selected that predominantly yield glucose as degradation product of
starch-based sizes. Amyloglucosidases performed best in this aspect. Bioscouring was
performed with combinations of enzymes that are promising in regard to both goals – to
significantly increase fiber absorbency and to yield sufficient glucose from the decomposition of
the non-cellulosic impurities. Pectinase, either as the only scouring compound or applied
together with other enzymes yielded good results. Lipases supported the water absorbency.
The addition of cellulase improved the hand of the textile, but caused a higher weight loss and
slight fiber peeling or cracks along the fiber axis as typically observed with cellulase finishing
procedures.
Glucose oxidases were the enzymes of choice for bio bleaching because; they use generated
glucose from starch-desizing and bioscouring for in-situ hydrogen peroxide production. It was
determined that generally 500-600 mg/L hydrogen peroxide were necessary for generating a
satisfactory whiteness level. After optimization of the separate steps regarding end-performance
of the treated goods, the steps were combined. The treatment baths of both scouring and desizing
were used separately at first and the content of glucose determined, then also combined. To
produce the necessary amount of peroxide, the available glucose and the dosage of the glucose
oxidase have to be carefully balanced. In this system, higher peroxide levels did not improve the
whiteness levels, possibly due to the stabilizing effect of the simultaneously produced gluconic
acid.
With bleaching at pH 7 and peroxide solely produced by enzymatic means, whiteness indices of
the treated goods of 62-64 could be achieved, which are close to those of conventionally scoured
and bleached cotton samples (WI = 72). Absorbency and mechanical properties of the
enzymatically treated goods were excellent and the fiber damage kept to a minimum.