Journal of Cleaner Production xxx (2013) 1e8

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Journal of Cleaner Production

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Enzyme mediated beam house operations of leather industry:

a needed step towards greener technology
Saurabh Saran a, Richi V. Mahajan b, Rekha Kaushik a, Jasmine Isar b, Rajendra K. Saxena a, *
a
b

Technology Based Incubator, University of Delhi South Campus, Benito Juarez Road, New Delhi 110021, India
Department of Microbiology, University of Delhi South Campus, Benito Juarez Road, New Delhi 110021, India

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 10 January 2013
Received in revised form
8 April 2013
Accepted 13 April 2013
Available online xxx

Leather industries, due to pollution of water, careless disposal of solid wastes and gaseous emissions, are
categorized as a red industry and are under deep pressure to develop environmentally efcient leathermaking processes to comply with modern pollution and discharge legislation. In the present study,
completely enzyme based beam house operations (dehairing & degreasing) of leather industry has been
standardized for skins & hides using Bacillus licheniformis protease and Bacillus subtilis lipase. Owing to its
immense importance and demand the productions of these enzymes were optimized and scaled up in a
300 L bioreactor resulting in a maximum yield of 4568 U/mL in 18 h for protease and 34.91 IU/mL of
lipase in 36 h. 100% enzymatic dehairing & degreasing of skins and hides could be obtained at pH value of
8.0 and temperature 30e37
C with the enzyme concentration of 2e5% (w/v) of protease for dehairing
and 5e10% (w/v) of lipase for degreasing within 8e12 h. An interesting observation noted for enzyme
based leather processing was that natural skin colour was preserved along with a drastic reduction of
BOD and COD values of efuent. Crust leather formed using enzymatically processed skins exhibited
similar physical and tactile properties as observed for conventional crust leather. On the bases of
internally quenched uorescent substrate specic method, this protease is subtilisin-like serine protease
with low collagenase activity. High collagenase activity can damage the hide (leather) grain and the
physical-mechanical characteristics of the hides. Successful demonstrations of large size (Industrial
trials 100 kg skin/hide at Central Leather Research Institute, Chennai, India) 100% chemical free
dehairing and degreasing further supports the authenticity of the work. Thus all this makes the beam
house operations of leather tannery enzymatically feasible and environmentally benign.
2013 Elsevier Ltd. All rights reserved.

Keywords:
Leather processing
Dehairing
Degreasing
Protease
Lipase

1. Introduction
The leather industry has already marked a place of prominence
in the Indian economy and national planning in view of its massive
potential for employment generation, growth and exports. This
industry employs more than 2.5 million people in the country, of
which 30% are women. Leather industry nds major applications
in garments, footwear, bags, carpets, tents, rugs etc. (Jegannathan
and Nielsen, 2013). According to the company ber2fashion
(18 February, 2011), there has been a remarkable rise of global trade
of raw leather from US $14.98 thousand million in 2000 has
reached to around US $20.63 thousand million in 2010. It is forecasted that this may rise to $32 thousand million by 2020. Current

* Corresponding author. Tel.: 91 (0)11 24116559; fax: 91 (0)11 24115270.

E-mail addresses: rksmicro@yahoo.co.in, rksmicro@hotmail.com (R.K. Saxena).

worldwide market of raw leather and leather goods are of $137

thousand million with the projection to touch $245 thousand
million by 2020 from the current level. (Fibre2fashionNews Desk).
The Indian leather industry has evolved over nearly two centuries.
There are about 2100 tanneries located in India, with a processing
capacity of 10  106 tons of raw hides and skins, making this a
thrust area in national planning for the development of India
(Vellappan and Muralidharan, 2001).
Conventional leather processing involves several unit operations, in which the use of chemicals like lime, sulde, chrome in
different stages of leather processing especially in pre-tanning and
tanning processes brings about almost 80e90% of the pollution
(Thanikaivelan et al., 2004). Besides this sulde, liberates toxic
hydrogen sulde, a serious hazard for both tannery workers and
sewer men (Arunachalam and Saritha, 2009). It makes leather
processing being an intensive source of producing large amounts of
liquid, solid and hazardous wastes (Saravanabhavan et al., 2005).

0959-6526/$ e see front matter 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jclepro.2013.04.017

Please cite this article in press as: Saran, S., et al., Enzyme mediated beam house operations of leather industry: a needed step towards greener
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S. Saran et al. / Journal of Cleaner Production xxx (2013) 1e8

Liming operation for the removal of hair and unwanted adhering

subcutaneous layer causes nearly 40% of biochemical oxygen demand (BOD) and 50% of chemical oxygen demand (COD). Hair as
pulp and subcutaneous layer is a major contributor to BOD while
COD is due to large amounts of chemicals used for dehairing (Rao
et al., 2002). Moreover, the use of lime in the unhairing process
requires its removal, usually by the addition of ammonium salts,
one of the main contaminants of liquid efuents (Dettmer et al.,
2012). Hence, alternatives to chemicals like sulphide, lime mediated dehairing and degreasing are being actively sought (Dayanand
et al., 2003). Conventional degreasing methods use large amount of
organic solvents and surfactants which give rise to environmental
problems such as volatile organic compounds (VOC) emissions.
Lipase can remove fats and grease from skins and hides, particularly
those with a moderate fat content (Hasan et al., 2010). Deliming
and bating are the most suitable processing places for using lipase
in the beam house operation in the leather industry.
Due to increased legislation geared towards environment protection and preservation of water qualities, tanneries are under
constant pressure to implement source reduction technologies that
reduce their organic and inorganic waste water loading (Tayler
et al., 1987). In this respect, developments of enzymatic processes
for dehairing and degreasing are being encouraged because they
not only yield quality improved products but also reduce use of
hazardous and polluting chemicals (de Souza and Gutterre, 2012).
In this context, protease and lipase enzymes seem to be quite
promising as they can be efciently used in the beam house operations of tanneries for leather manufacturing processes. The present work modies the conventional process and employs lime
free, hundred percent efcient enzymatic dehairing and degreasing
of goat skins and buffalo hides. Here, the enzymes used for
dehairing (protease) and degreasing (lipase) experiments were
produced in a 300 L fermenter and further optimization was undertaken to achieve complete beam house operations of leather
industry.
2. Materials and methods
2.1. Organism and growth conditions
Bacillus spp., potent protease (SBP-114) and lipase (RK-2) producers were isolated from the soil by enrichment and selective
screening on skim milk agar plate and tributyrin agar plate
respectively. These strains were identied by MIDILABS (New York,
DE, USA) as Bacillus licheniformis and B. subtilus based on 500-bp
analysis and on the partial 16S rRNA gene alignment with the Gene
Bank database (ncbi, 2012) (). The organisms were cultivated at
37  1
C in a bacteriological incubator for 24 h and subsequently
maintained at 4
C in a B.O.D incubator (Yorko, Deluxe-10) by
routine transfers on nutrient agar slants at pH value of 7.0.
2.2. Enzyme production in a bioreactor
The enzymes used for dehairing (protease) and degreasing
(lipase) were produced and optimized in a 300 L stirred bioreactor
(Scigenics, India) with a working volume of 220 L, separately. The
modied base medium (pH-7) used for protease production contained (gL1): wheat bran (2); soybean meal (15); KH2PO4 (1.0);
K2HPO4 (3.0); Na2SO4 (2.0); and MgSO4.7H2O (0.10). The medium
used for lipase production contained (g/L): sunower oil (2);
glucose (2); peptone (15); NH4NO3 (3.0); and MgSO4.7H2O
(0.8 mM) (pH 8). These media were sterilized separately in situ at
121
C for 20 min and inoculated with 2.0% of the seed inoculum
(nutrient broth) (OD660nm y 0.600). Fermentation for protease
production was carried out at 37  1
C for 24 h and for lipase

30  1
C for 36 h. Rest of the parameters remained same for both

the enzyme production. The impeller speed was initially adjusted to
350 rpm and compressed sterile air was sparged into the medium at
constant rate of 0.6-VVM. The dissolved oxygen was not allowed to
fall below a xed set point of 40% by cascading in both the cases.
Samples were withdrawn periodically at 2 h intervals and analysed for protease and lipase production. The fermentation parameters, such as temperature, pH, dissolved oxygen and airow
were continuously monitored using microprocessor-controlled
probes. The fermentation broth was centrifuged at 8000 g for
20 min at 4
C to obtain cell free broth, containing lipase and
protease respectively. The cell pellet was discarded and the crude
enzyme was used for dehairing and degreasing experiments.
The crude protease and lipase rich broth were further concentrated four times separately by ultraltration (using Spintrex) using
10 kDa MWCO membrane cartridge.
2.2.1. Protease assay
The protease activity was measured by the procedure as
described by Meyers and Ahearn (1977) with some modications
(Saran et al., 2007). One unit of protease was equivalent to the
amount of enzyme required to release 1 mg of tyrosine per mL per
minute under standard assay conditions.
2.2.2. Lipase assay
Lipase activity in the culture ltrate was determined spectrophotometrically using p-nitrophenyl palmitate as the substrate
using the procedure of Winkler and Stuckman (1979). One International Unit (IU) of lipase activity was dened as the amount of
enzyme required to release 1 mM of p-nitro phenol per mL per
minute under the standard assay conditions.
2.3. Internally quenched uorescent substrate specic method
In order to further conrm that B. licheniformis protease is
subtilisin-like serine protease with low-collagenase activity, the
concentrated protease samples were evaluated by internally
quenched uorescent method. This method is based on the use of
designed, internally quenched uorescent universal protease
substrates, which contain several specic cleavage sites. Application of a combination of uorescence and mass spectral analysis
permits the proling of proteolysis.
2.4. Dehairing & degreasing of animal skins and hides
2.4.1. Preparation of animal skins and hides
Goat skin, sheep skin and buffalo hide were obtained from the
local slaughter house. For storage, the skins were rubbed with twice
their weight in salts and stored in cold room at 4
C for 2e6 months
to prevent putrefaction. The skins were cut to a size of 1 sq. ft
(0.093 m2) for experiments. The salted skins and hides were
soaked, for a period of 6 h in water to eliminate salt, blood, and dirt
and to gain moisture lost during curing process.
2.4.2. Experimental
The paste method was followed in all the studies for the beam
house operations of the leather industry (Fig. 4A,B). The skins/hides
after application of enzymes were loosely covered with single layer
of polythene sheets and kept at room temperature (30  2
C) for
24 h. Dehairing and degreasing were checked periodically up to
24 h. To check the dehairing potential, hair was removed using a
blunt end spatula. The efcacy of the dehairing enzyme at different
parameters (temperatures, pH, and enzyme concentrations)
was optimized for 100% dehairing. The scale of dehairing was
observed as follows: a) slight dehairing, b) moderate

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S. Saran et al. / Journal of Cleaner Production xxx (2013) 1e8

Fig. 1. Fermentation prole of protease in a 300 L fermentor.

dehairing and c) complete dehairing. These dehaired skins

were processed to crust leather.
The potency of lipase was checked by the weight loss in the skin
(removal of esh from the skin by spatula). The amount of fat
removed was calculated by subtracting the weight of treated skins
from its initial weight taken before treatment.

TDS, BOD, and COD. Sulde and calcium content in the efuent of
experimental group were not estimated since the developed process was free of sulde and lime.
3. Results
3.1. Production of enzyme in a 300 L bioreactor

2.4.4. Analysis of efuent after leather processing using enzyme

The efuent after leather processing by chemical based and
enzyme processed were analysed for various parameters such as

Several fermentor runs were carried out to optimize the production of protease and lipase in a 300 L bioreactor, separately.
Parameters such as agitation rate, dissolved oxygen and inoculum
levels were optimized. Final yields for protease production showed
that the inoculum level (3%), dissolved oxygen levels greater than
40% saturation rate, agitation rate 350 rpm and aeration 1.5 VVM
resulted in 4586 U/mL of protease in 18 h (Fig. 1). Optimization of
various physiological factors revealed that the major controlling
factor for lipase production were inoculums density (3.0%), agitation rate (250 rpm) and dissolved oxygen (D.O) levels at 40% saturations. The lipase production from the present strain of B. subtilis

35

12

25
4
20
3
15
2
10
1

0
6

12

18

24

30

36

42

48

Specific activity (IU/mg)

Lipase Production (IU/ml)

30

14

10

Growth O.D.660

2.4.3. Evaluation of physical properties of leather

Tensile and tear strength tests were carried out for the crust
leathers both (chemical & enzyme processed) along and across
backbone line. The mean of the values corresponding to along and
across backbone line was calculated for each skin for each strength
character. Physical properties such as tensile strength, percent
elongation at break, tear strength and grain crack strength were
also examined.

54

Incubation Period (h)

Fig. 2. Fermentation prole of lipase in a 300 L fermentor.

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S. Saran et al. / Journal of Cleaner Production xxx (2013) 1e8

industry because collagenase activity can damage the hide (leather)

grain and the physical-mechanical characteristics of the hides.
3.3. Preliminary trials of leather processing using enzyme

Fig. 3. Sequence of the universal protease substrate 1 and 2. Potential protease

cleavage sites are indicated.

could successfully be scaled up to a pilot plant level with a substantial yield of 34.9 IU/mL in 36 h (Fig. 2).

3.2. Internally quenched uorescent substrate specic method

This method is based on the principle of using universal substrate which represents a highly sensitive way of monitoring multiple proteases of varied specicities. This provides information
regarding the nature of the cleavage sequence preferred by individual enzymes and the design of the protease substrate showing
appropriate cleavage site for different classes of proteases, as analysed by MALDI eTOF (Pattanaik et al., 2003). If peptide 1 is cleaved
after Ala 10 e Ala 11 the enzyme is keratinase elastase like and if it
is cleaved after Ala 11 e Ala12 the enzyme is subtilisin protease K
like (Fig. 3). When this B. licheniformis protease was treated with
this substrate, the fragments obtained are Dab-Gaba-Arg-Pro-LeuGly-Ala-Ala-Ala- by uorimetry and MALDIeTOF-Mass Spectrometry. This fragment conrms that the present protease is
subtilisin- like serine protease with low collagenase activity
(Table 1). High collagenase activity is not desirable for leather

Preliminary trials were performed to evaluate the dehairing

potential of the enzyme (protease). Experimentally, 3 sets of
experiment were performed in which leather processing were
evaluated using (1) chemical method (chemical and water) (2)
enzyme assisted (enzyme and chemical) (3) enzyme mediated
(enzyme and water). It was observed that slight () to moderate
() deharing were observed when only crude enzymes (unconcentrated) were used (Table 2) in 30 h. However, complete
dehairing was observed with chemical and enzyme assisted
dehairing in 30 h.
Owing to potential of these enzymes (viz. protease and lipase),
the processes were standardized for the complete () dehairing
and degreasing of skin and hides. Here for efcient beam house
operations both lipase and protease enzyme were efciently
concentrated four times using ultra-ltration with 10 kDa membrane cartridge which resulted in 16,250 U/mL of protease and
142 IU/mL of lipase.
3.3.1. Standardization of conditions for efcient dehairing
Complete dehairing processes were standardized and optimized using concentrated B. licheniformis protease which resulted
in 2% (w/v) for goat skin, 1% (w/v) for sheep skin and 5% (w/v) for
the buffalo hide gave complete dehairing within 12 h (Table 3,
Fig. 4C, D). Efcient dehairing was obtained at pH value of 8 and 9
in 8 h. However, at pH value of 10 the period of dehairing
increased to 12 h or even more. At pH value of 7.0, hundred
percent dehairing was not observed even after 24 h of incubation.
The original pH of the concentrated protease is around 8.0, and
since complete dehairing was observed at pH value of 8.0, thus
there was no need of addition of any NaOH for pH adjustment.
At 30
C and above temperatures, 100% dehairing was observed in
8e10 h in sheep and goat skin and 10e12 h for buffalo hide. Even
though 100% dehaing was observed at 25
C, the incubation time
was much longer up to 24 h. Thus, this protease could also be
efciently used for dehairing in tanneries in cold countries where

Table 1
Fluorescence and mass spectral analysis of crude protease extract.
Name of extract

Protease (SBP-114)

Activity as measured
by uorescence*

Fragments obtained by MALDI-TOF

Observed (Da)

Expected

Fragment (Da) MH

Comment

920.6 (MH)
991.7 (MH)

918
989

Dab-Gaba-Arg-Pro-Leu-Gly-Ala-Ala
Dab-Gaba-Arg-Pro-Leu-Gly-Ala-Ala-Ala-

Subtilisin like eserine protease

with low collagenase activity

Table 2
Preliminary trials to evaluate protease enzyme potentials for dehairing of skins and hide and its comparative analysis with chemical, enzyme assisted leather processing.
Time (h)

8
12
16
20
24
30

Chemical dehairing

Enzyme assisted dehairing

Enzyme mediated dehairing

Set 1 (Lime 10% Sulde 2% Water 15%)

Set 2 (Enz. 2.0% Sulde1% Lime 5% Water 15%)

Set 3 (Enz. 10% Water 90%)

GS

SS

BH

GS

SS

BH

GS

SS

BH

e
e
e

e
e
e
e

e
e
e

e
e
e
e
e

GS: Goat skin; SS: sheep skin; BH: Buffalo hide; (): No dehairing; (): Slight dehairing; (): Moderate Dehairing; (): Complete dehairing, Enzyme activity: 4568 U/ml
(protease); 34 IU/ml (lipase).

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S. Saran et al. / Journal of Cleaner Production xxx (2013) 1e8

Table 3
Standardized conditions for efcient leather processing.
Optimized conditions

Enzyme concentration
pH
Temperature
Incubation period

Goat skin

Sheep skin

Buffalo hide

Dehairing (100%)

Deeshing
(>95% fat removal)

Dehairing (100%)

Deeshing
(>95% fat removal)

Dehairing (100%)

Deeshing
(>95% fat removal)

2%
8
30e37
C
8e10 h

8%
8

1%
8
30e37
C
6e8 h

5%
8

5%
8
30e37
C
10e12 h

10%
8

12 h

the ambient temperature is around 25
C. This also is an effort to

make the beam house operation of the tannery completely green
without the assistance of any chemicals.
3.3.2. Standardization of conditions for efcient degreasing
Bacillus subtilis lipase carried out successful degreasing (>95%)
of goat and sheep skin and buffalo hides with 5e10% lipase in 12 h
of application without the use of any chemicals (Table 3, Fig. 4E, F).
Concentrated lipase having 142 IU/mL when applied on the esh
side for approximately 12 h, there was a noticeable decrease in
weight of the goat, sheep skins and buffalo hide resulting in 961,
926 and 914 g from the initial weight of 995 g, 997 g and 993 g
respectively. This indicates successful removal of 34, 67 and 83 g
of fat from the goat, buffalo and sheep skins respectively after
treatment with this lipase.

10 h

12e14 h

3.4. Evaluation of physical properties of leather

The results between the chemical and enzyme processed
leathers were compared. It was found that the enzyme processed
leather has a better tensile strength (220 kg/cm2), better percentage elongation (81%) and tear strength (71 kg/cm2) as compared to
the chemical processed leather, (Table 4).
The strength properties of the leather obtained from the
enzyme mediated process were better compared with conventionally (chemical) processed leathers. Fullness, grain smoothness and grain tightness of the leather from the enzyme
mediated process are also better than the conventionally processed leather. Softness and general appearances of the leather
from the rationalized process are comparable to the conventionally processed leather (Fig. 5A). In addition to this, the skin

Fig. 4. Dehairing and degreasing of goat skin after enzyme treatment.

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S. Saran et al. / Journal of Cleaner Production xxx (2013) 1e8

Table 4
Physical testing data of chemically processed (C) and enzyme processed (E) leather.
Sample

C
E
UNIDO norms

Tensile strength (kg/cm2)

185  5
220  5
120

Percent elongation

80  4
81  3
e

Tear strength (kg/cm2)

63  4
71  4
20

Grain crack strength

Bursting strength

Load (kg)

Distension (mm)

Load (kg)

Distension (mm)

24  1
30  2
e

11  0.4
14  0.5

65  1
64  2
e

16  0.4
15  0.5

The report values are average of ve measurements along with standard error.

had a much cleaner look with no shrinkage (original size)

retaining its original colour and sheen (Fig. 5B). The overall
performance of both chemical and enzyme based leathers are
comparable.

BOD and COD for enzymatic processing of buffalo hides was 82%
and 85%, respectively (Table 5).

3.5. Analysis of efuent after enzyme processed beam house

operation

Industrial trials of the beam house operations of leather industry were carried out at the tannery Central Leather Research
Institute (CLRI), Chennai, India using 100 kg skin/hide. Complete
dehairing and degreasing were observed under the tannery conditions using these protease and lipase. The goat skins and buffalo
hides retained their original shape, size, texture and colour. The
hairs were removed right from the hair follicle and were intact
proving that it was possible to use the present protease for

A reduction of around 86% TDS load in the efuent from the

enzymatic processing of skins and hides against the conventional
process. BOD and COD load from the efuent of enzymatic processing of goat skins were reduced by 75% and 80%, respectively
when compared to conventional processing. Similarly, reduction of

3.6. Industrial trials of enzyme mediated dehairing

Fig. 5. (A): Comparison of functional properties of leather from chemically processed and enzyme processed leather. (B): Comparative analysis of chemical processed and enzymatic
processed skin. (C): Microscopic view of an intact hair. (D): pH prole obtained after chemical (red) and enzymatic dehairing (green). (For interpretation of the references to colour
in this gure legend, the reader is referred to the web version of this article.)

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Table 5
Analyses of pollution parameters and load from the efuent of dehairing processes
of skin and hide.
Parameters efuent
load (kg/t)

Goat skin
Conventional

Enzymatic

Conventional

Buffalo hide
Enzymatic

TDS
BOD
COD
Ca2
Sulde

19.45
3.84
12.35
2.10
3.12

2.81
0.95
2.42
e
e

28.54
9.24
27.34
2.31
4.22

5.13
1.65
4.10
e
e

dehairing and lipase for degreasing process in beam house operations without use of any chemicals.
4. Discussions
Maximum protease production was observed at the end of
exponential phase. Thereafter, a decline in the protease production
was observed in all the fermentors. Gupta et al. (2002) have also
reported a similar cessation in protease production once a
maximum amount of the enzyme is produced during the run. Our
results are further supported by Genckal and Tari (2006) who
while working with Bacillus sp. and and Potumarthi et al. (2007),
who while working with B. licheniformis NCIM-2042 reported a
decline in protease production after stationary phase. It has been
observed that mostly maximum lipase yield is achieved at a time
when the lipid (oil/fat) is completely consumed by an organism in
the growth medium (Li and Zhang, 2005). In the present study
similar ndings have been observed, where in 36 h oil was
consumed and maximum lipase was produced. In the present
study, a maximum of 34.9 IU/mL of the enzyme was obtained in
36 h at 2.0% inoculum.
The pre-tanning operations alone contribute to 70e80% pollution loads in tannery waste water (Dayanand et al., 2003). Conventional leather-processing methods subject the skin or hide to
wide variations in pH (Ludvik, 1977). During the pre-tanning
operation pH of the skin is changed from neutral pH (soaking) to
highly alkaline pH value of 12e13 (liming). In this operation, lime
and sulde are used with substantial quantities of water. Again
during deliming processes ammonium chloride and ammonium
sulphate are used to neutralize the pH. Further during bating and
pickling processes the pH of the skin is adjusted to pH value of
3.5e4.5 and further 2.5e2.8 respectively, using sulfuric acid and
sodium chloride. Subsequently, during the time of tanning and
post tanning pH value increased to 6.0. Such changes in pH demands the use of acids and alkalis, which lead to the generation of
salts, resulting in a net increase in chemical oxygen demand (COD),
total solids (TS), chlorides (Cl-) and sulphates (SO2
4 ) in tannery
waste water (Thanikaivelan et al., 2004). Thus, the wide variations
in pH during the various steps of leather making are also bound to
impair the bulk as well as the surface properties of hide/skin
matrix (Huber and Satyendra, 1990). However, when the skin and
hides were processed using enzymatic approach, liming, deliming,
and bating process could successfully be carried out at pH value of
around 8.0 (Fig. 5C). The enzymatic tanning process does not
require sodium chloride and sulfuric acid. Thus, the total amount
of chemical and byproducts consumed in the conventional processing is around 385 and 54 kg, respectively. This means that
enzymatic leather processing enjoys a reduction in total chemical
consumption of 85% compared to chemical leather processing.
Kandasamy et al. (2012) also reported enzyme-dehaired leathers
exhibit similar or improved characteristics. Also, the signicant
decrease in processing time, water and chemical consumption

during enzymatic leather processing has been reported by Olle

et al. (2013). In chemical based dehairing and deeshing, the
hairs uprooted from the skin get broken into small pieces, thereby
increasing the BOD, COD, TDS of the water and the skin is also
shrunken and distorted in shape. This also results in an unclean
and dull grey appearance of the dehaired skin and hide. However,
when the skins and hides were processed using enzymes, the
protease opens up the membrane surroundings of the fat cell,
making the fat accessible to the lipase and it was observed that the
hair comes out of the hair follicle unbroken and completely
uprooted from the hair follicle (Fig. 5D). Krishnamoorthy et al.
(2013) reported that these hairs can further be used in the making of fur jackets, cap, globes, etc.
5. Conclusions
The present investigation is a breakthrough approach in making
the beam house operations of the leather chemical free. It was a
successful attempt to modify the conventional chemical process to
a lime free, environmentally benign technology. The productions of
enzymes were successfully scaled-up in 300 L stirred reactor
resulting in a maximum yield of 4568 U/mL in 18 h for protease and
34.91 IU/mL of lipase in 36 h. Hundred percent efcient enzymatic
dehairing and degreasing of skins and hides were achieved at pH
value of 8.0 and temperature 30e37
C with the enzyme concentration of 2e5% (w/v) of protease (B. licheniformis) for dehairing and
5e10% (w/v) of lipase (B. subtilis) for degreasing within 8e12 h of
incubation. Enzymatic leather processing also signicantly decreases the processing time, BOD, COD, water and chemical consumption during the operation. The bulk size tannery trials at CLRI,
Chennai were efcient and successful leading a way for changing
the beam house operation from chemical to enzyme assisted and
now nally to total enzyme based leather processing. This all forms
a part of making the tannery go green.
Acknowledgements
The authors thank Technology Based Incubator, UDSC, New Delhi
for providing the infrastructure support. Dr. Saran thanks New Millennium Technology Leadership Initiative (NMITLI) of CSIR for
providing the nancial support to carry out the research. Sincere
thanks to Prof. P. Balram, Director, IISC, Bangalore for protease
sample analysis by Fluorescence and Mass spectral analysis. The help
and support of Dr. Ramasami, (secretary, DST) the than Director,
Centre Leather Research Institute (CLRI), Chennai for allowing us to
conduct industrial trails at CLRI is also deeply acknowledged.
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Further reading
HYPERLINK "http://<www.bre2fashion.com/news/leathers-news/newsdetails.aspx?
news_id96002>" \o "http://<www.bre2fashion.com/news/leathers-news/
newsdetails.aspx?news_id96002>"<www.bre2fashion.com/news/leathersnews/newsdetails.aspx?news_id96002> dor. February 18, 2011.
HYPERLINK "http://<www.ncbi.nlm.gov./BLAST./blast _help. htm>" \o "http://
<www.ncbi.nlm.gov./BLAST./blast _help. htm>"<www.ncbi.nlm.gov./BLAST./
blast _help. htm> dor. May 26, 2012.

Please cite this article in press as: Saran, S., et al., Enzyme mediated beam house operations of leather industry: a needed step towards greener
technology, Journal of Cleaner Production (2013), http://dx.doi.org/10.1016/j.jclepro.2013.04.017