Crude-Protease-Based Green Leather: Unhairing Technology for

Enhanced Properties and Pollution Reduction

Md. Jawad Hasan1, Md. Samaul Alam1, Papia Haque2, Mohammed Mizanur Rahman1,2
1Institute
of Leather Engineering and Technology, University of Dhaka
2Department of Applied Chemistry and Chemical Engineering, University of Dhaka

Abstract Results
Traditional beam house chemicals like lime and sodium sulfide, used to remove hair from hide/skin and
open skin fibers during leather processing, cause excessive water consumption, hair digestion, and solid
sludge discharge into the effluent. Enzyme-only unhairing and fiber opening have been introduced as a
solution to address the environmental concerns associated with conventional chemical unhairing.
Proteolytic bacteria from Alcaligenaceae sp. were isolated from the soil of tannery solid waste (TSW)
dumping yard. The isolated bacteria produced protease with high activity at 50°C and pH 7.0. The crude
protease was used to completely unhair goatskin and increase fiber opening, replacing three highly
polluting stages (liming, deliming, and bating) from conventional leather processing practice. The use of
protease-based unhairing resulted in significant pollution load reduction in terms of dissolved solids,
suspended solids, and total solids compared to conventional liming. The crust leathers produced from
protease-based unhairing showed better hydrothermal stability, and physical strength properties.
Microscopic imaging confirmed the fiber structure of the experimental sample being more compact than the
conventional leather. A clean, unhairing procedure using Alcaligenaceae sp. crude protease can make
"green leather" without the use of sodium sulfide, lime, ammonium-based deliming agents, or bating agents.

• 97.06% similarity with Alcaligenaceae at • 10% inoculum size was sufficient for
family level. maximum protease production
• 24 hours incubation time period for
maximum protease production in Media 3

29.57 U/ml 35.54 U/ml

Figure: Some of the specific purposes of unit processes of leather making • The obtained crude protease showed highest activity at 50°C temperature and at pH 7.0.
Table: Pollution load by unhairing in terms of TDS, TSS and TS.
Sample TDS Removal TSS Removal TS Removal EC Removal
(weight (mg/L) (%) (mg/L) (%) (mg/L) (%) (mS/cm) (%)
Methodology ratio)
1 (1:1) 20700±152.7 20710.32±151.3
5 27.7 44±1.51 89.27 5 28.56 41.4±1.35 27.75
2 (1:1.5) 18950±255.8 33.9 32±1.3 92.20 18688.5±255.8 35.53 37.9±2.03 33.86
3 (1:2) 15650±446.8 45.4 50±1.27 87.80 15628.16±445.6 46.09 31.3±1.15 45.38
4 (1:2.5) 13950±544.1 51.3 8.98±0.6 98.05 13659.28±543.5 52.88 27.9±1.13 51.31
5 (1:3) 11122.07±272.6
11425±272.3 60.1 6±0.5 98.54 3 61.63 22.85±0.96 60.12
Conventiona 28650±503.1 - - - -
l Liming 4 410±2.5 28990±500.7 57.3±1.26

First crude Rough unhairing

Proteolytic bacteria isolation protease trial for Production of
production confirmation crude protease

Histology and Second level First level Activity

application application measurement of
Pollution load test
crude protease
Serial no. Amount of Amount of
crude crude protease
protease used used in the
in the first second step
step
Complete Sample 1 50% 100%
leather Sample 2 75% 150%
making Sample 3 100% 200%
Sample 4 150% 250%
Sample 5 200% 300%
Sample 6 250% -
Sample 7 300% -
Sample 8 400% -
Sample 9 500% -
Table: Physical strength properties of leathers produced from enzymatic unhairing and
Characterization of crust conventional liming.
leathers
Sample Tensile Elongation at Stitch tear Grain Distension Shrinkage Total Cr
• Chrome uptake test (weight ratio) strength break (%) strength crack (mm) temperature %*
• Shrinkage temperature (kg/cm2) (kg/cm) load (kg) (°C)
• Strength properties 1 (1:1) 223.31 93.91 110.21 9.48 29.0 103.0±1 1.06
• FTIR and TGA analysis 2 (1:1.5) 232.87 67.15 115.45 10.73 35.0 102.5±2 1.04
• FESEM imaging 3 (1:2) 235.65 60.95 121.31 10.04 28.0 102.0±1 1.32
4 (1:2.5) 238.41 79.34 132.15 10.39 25.0 101.0±1 1.27
5 (1:3) 232.34 89.25 118.68 10.86 32.0 105.0±2 1.17
Conventional 160.64 99.87 97.58 8.96 20.0 102.5±1 1.05
Liming
Minimum 200 40-60 60-80 20 7.00
Standard**
UNIDO 120 - - - -
Standard***
. *Based on the dry weight of the wet blue leather sample; **(Kanagaraj et al., 2001); ***(Christopher et al.,
2014).

Conclusion
This study highlights proteases as an eco-friendly alternative in leather processing. Using proteases from locally isolated bacteria
like Alcaligenaceae sp., with high activity at 50°C and pH 7.0, it demonstrates cleaner leather production with reduced
environmental impact. This approach enhances leather quality, meets industry standards, and improves chromium absorption,
Acknowledgements offering ecological and economic benefits. Protease-based unhairing produces crust leathers with enhanced stability and
strength. Microscopic imaging confirms a denser fiber structure compared to conventional leather. These findings suggest
proteases can revolutionize leather production sustainably. Promoting their adoption, optimizing cost-effectiveness, and exploring
Professor Dr. Abu Ashfaqur Sajib wider applications are crucial for advancing sustainable leather production.

Chairman, Department of Genetic Engineering and Biotechnology, University of Dhaka