Journal of Microscopy Society of Thailand 2009, 23(1): 142-146

Papain Enzymatic Degumming of

Thai Bombyx mori Silk Fibers
Monthon Nakpathom*, Buppha Somboon, Nootsara Narumol
National Metal and Materials Technology Center, National Science and Technology Development Agency,
Thailand Science Park, Phathumthani, Thailand
*Corresponding author, e-mail: monthonn@mtec.or.th

Abstract
Thai Bombi mori silk fibers were degummed with a papain enzyme derived from local papyrus latex and
compared with a traditional alkali/soap method. The efficiency of degumming processes was assessed using
percent weight loss, tensile strength and scanning electron microscopy (SEM) measurements, color staining
testing and dyeability with a natural lac dye. The percents weight loss of papain enzymatic and alkali/soap
degummed fibers were not significantly different, i.e., 20-22%. The decreases in tensile strength were 46%,
40.2% and 28% for silk fiber degummed with 5% owf papain enzyme at high temperature (65-70 C for 1 h),
alkali/soap and 10% owf papain enzyme at room temperature for 24 h, respectively. The surface morphological
of the degummed fibers by SEM analysis confirmed the removal of sericin from silk fibroin. Higher color yields
(K/S values) were obtained in case of lac dyeing of silk fibers degummed with papain enzyme with having fair
to good color fastness to washing and light.

Papain is a proteolytic enzyme produced from

the latex of the green papaya fruits [7]. It has high
protein hydrolyzing capacity. Major applications of
papain enzyme include food industries, primarily
for chill proofing of beer, tenderizing of meat and
freeing of food proteins. Other applications are in
tanning of leather and hides, degumming of silk,
cheese manufacture, pharmaceuticals, aroma and
perfume industries and effluent treatment.
The aim of the present research is to investigate
the influence of papain enzymatic degumming of
Thai Bombyx mori silk fibers on fiber damage and
residual sericin by percent weight loss calculation,
tensile strength and scanning electron microscopy
(SEM) measurement and color staining testing. The
traditional alkali/soap degumming method was also
performed for comparison. In additions, the
resultant degummed silk fibers were dyed with a
natural lac dye in order to evaluate their dyeability.

Background
Thai Bombyx mori silk, produced by cultivated
mulberry silkworm, is greatly appreciated as one of
the most luxurious textile commodities. Known for
its unique characteristics such as luster and
softness, the production of Thai Bombyx mori silk
consists of silkworm raising, cocoon production,
silk reeling, degumming, dyeing, weaving, and
finishing products [1]. Degumming is the process
where sericin, i.e., the silk gum gluing the fibroin
filaments, is totally removed in order to obtain a
typical shiny, soft handle and other desirable
properties [2]. Due to its highly oriented and
crystalline structure, fibroin is not water soluble,
whereas sericin is easily dissolved in a boiling
aqueous solution with the presence of soap, alkali,
organic acid or synthetic detergents. Nowadays,
batch degumming of silk is mostly carried out in an
alkaline bath with pH about 10 containing 10-20
g/L soap at 92-98 C for 2-4 h [2-3]. During highly
hot alkaline silk degumming, hydrolytic
degradation of fibers is accompanied, leading to
undesirable both aesthetic and physical properties
such as dull appearance, surface fibrillation and
tensile strength loss. Thus, synthetic detergents
may be used to replace soaps in case of reduced
treatment time and less fiber damage are needed.
Furthermore, enzymatic treatment of silk is
currently gained large attention as an alternative for
degumming process owing to its ecologically safe
and required milder conditions [4-6]. Proteases and
lipases are normally used in combination for
degumming and removing others impurities such as
waxes, fats, mineral salts and pigments [2-3].

Materials and Methods

Materials
The Bombyx mori silk fiber used in the
experiment was sourced from the northeast part of
Thailand. Papain enzyme was derived from
papyrus latex of Thais green papaya fruits with
1.5-3 months of age. The collected latex was
successively dried in the open air and in the oven at
60 C and then ground into a yellowish powder.
Lac dye was obtained from the northeast of
Thailand. Alum (AlK(SO4)2.12H2O) was purchased
from Asia Pacific Specialty Chemicals Ltd.,
Thailand. All other chemical reagents were of
analytical grade and purchased from either Aldrich
Chemical Company or Merck Ltd.

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Journal of Microscopy Society of Thailand 2009, 23(1): 142-146

dyed with a lac dye solution in a liquor ratio of

1:30 at 90-95 C for 45 min. Dyed silks were then
rinsed with water and air-dried.
Color measurement and testing
The color strength (K/S values) and CIELAB
(L*a*b*) values of dyed silk fibers were measured
on the Gretag-Macbeth Color i5 spectrophotometer
with illuminant D65 at 10 degree observer. Color
fastness tests to washing and light were determined
according to ISO 105-C01 (1989) and AATCC
Test Method 16 (2004), using DaeLim Starlet II IR
lab dyeing machine and Suntest CPS testing
equipment, respectively [10-11].

Silk degumming with alkali/soap

Silk samples were treated with an alkaline bath
containing an aqueous solution of 10 g/L soap and
2 g/L sodium carbonate in a liquor ratio of 1:30 at
90-95 C for 45 min. Degummed silks were then
washed with hot and cold distilled water and finally
air dried.
Silk degumming with papain enzyme
In our preliminary experiment, it was found that
both papain enzyme concentration and condition
(temperature and time) played crucial roles in order
to remove sericin from the silk fibers. In addition,
the optimum concentrations of papain enzyme were
at 5% and 10% owf were required for degumming
at 65-70 C for 1 h and at room temperature for 24
h, respectively. Hence, in this work, silk samples
were degummed according to the above deduction.
Degummed silks were then washed with hot and
cold distilled water and finally air dried.
Determination of the weight loss
The percent weight loss of the silk fibers after
degumming process was calculated from Eqn 1:
W 0 -Wt
% Weight loss =
100
W0

Results and Discussion

Table 1 summarizes the results on efficiency of
alkaline/soap and papain enzymatic degumming
processes of silk fibers in terms of percent weight
loss, breaking load and color staining. Although
there was a similar reduction in percent weight loss
(i.e., approximately 20-22%) of Bombyx mori silk
fibers degummed with alkali/soap and papain
enzyme, a distinct difference in breaking load was
clearly observed. Degumming silk fibers with 5%
owf papain enzyme at high temperature (65-70 C
for 1 h) yielded a maximum decrease of breaking
load, i.e., about 46% of the raw silk. On the other
hand, the breaking loads of silk fibers degummed
with 10% owf papain enzyme at room temperature
for 24 h and with alkali/soap dropped to 28.4% and
40.2%, respectively. This was probably due to an
intense enzyme activity at a higher temperature
condition compared to at room temperature.

(1)

where W0 and Wt are the weight of the silk fiber

samples before and after degumming, respectively.
Determination of breaking load
Breaking loads of silk fibers were performed
according to ASTM D 3822-01 Standard Test
Method for Tensile Properties of Single Textile
Fibers [8]. A single fiber was measured at a gauge
length of 250 mm and extension rate of 30 mm/min
on a tensile strength tester (LLOYD Instruments
LR5K, Intro Enterprise Co., Ltd.).
Scanning electronic microscopy (SEM) analysis
Morphological characterization of silk fibers
was performed by means of scanning electron
microscopy (Model 5410, JEOL). Samples were
observed at 15 kV acceleration voltage after gold
sputtering.
Color staining test
Ambidirect Red 3BL 165% (C.I. Direct Red
80), supplied by Thai Ambica Chemicals Co., Ltd
was used to evaluate the remaining of sericine after
silk degumming. The degummed silk samples were
immersed in an aqueous solution of 1 g/L
Ambidirect Red 3BL 165% with a liquor ratio of
1:200 for 2 min at the boil [9].
Lac dyeing
Crushed stick lac was first extracted by water
(weight ratio = 1:10) at the boil for 1 h and then
filtered to give a red solution of lac dye. The lac
dye solution showed a maximum absorption peak
(max) at approximately 485 nm. The degummed
silk fibers were pre-mordanted with 10% owf alum
solution at room temperature for 30 min and then

Table 1 Percent weight loss, breaking load and color

staining results of Bombyx mori silk fibers.

Degumming
Methods

Weight
loss (%)

Breaking
load (N)

Staining
Test

Raw silk

8.0540

Dark red

4.8190

Pale pink

4.3475

Pale pink

5.7632

Pale pink

Alkaline
21.94
/Soap1
Papain
20.60
5 %owf 2
Papain
20.80
10 %owf 3
1
90-95 C, 45 min
2
65-70 C, 1 h
3
room temperature, 24 h

For the direct dye staining test, raw silk was

stained dark red color as a result of a large quantity
of sericin present in the fibers. Both papain and
alkali/soap degummed silk appeared pale pink,
indicating that there were a small amount of sericin
remaining. Moreover, dull and stiff handle of raw
silk fibers disappeared and the degummed silk
became more shiny and softer.

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(a)

(c)

(b)

(d)
Fig. 1 SEM images of the silk fiber surface before and after degumming: (a) before degumming, (b) after degumming
with alkai/soap, (c) after degumming with 5%owf papain enzyme at 65-70 C for 1 h, (d) after degumming with
10%owf papain enzyme at room temperature for 24 h.

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Table 2 Color strength (K/S values), CIELAB L*a*b* values and color fastness of degummed silk fibers dyed with a
natural lac dye

Degumming
methods

Dyed
samples

CIELAB

K/S

Color fastness

L*

a*

b*

Washing

Light

Alkaline
/Soap

6.17

44.65

38.78

1.39

Papain
5 %owf

8.72

37.28

35.29

0.64

2-3

2-3

Papain
10 %owf

9.78

35.86

34.23

1.34

2-3

L = Lightness (0 = black, 100 = white)

a* = Red-green coordinate (positive values = red, negative values = green)
b* = Yellow-blue coordinate (positive values = yellow, negative values = blue)
The rating scale of color fastness to washing and light is from 1 (poor) to 5 (excellent)

natural dyes could be also undertaken as another

assessment of the efficiency of papain enzymatic
degumming.

SEM observation reveals that the raw Bombyx

mori silk fibers were covered with a relatively large
amount of sericin on fibroin as illustrated in Figure
1. Degumming of the fibers with alkali/soap and
papain enzyme at 65-70 C or at room temperature
could almost completely remove sericin. However,
fibrillation of some fibroin threads occurred during
the degumming processes.
In view of natural lac dyeing, papain enzymatic
degummed silk fibers had higher color strength
(K/S values) than those degummed with alkali/soap
as shown in Table 2. Meanwhile, there was no
significant difference in color fastness to washing
and light among these three processes. All dyed
silk fibers exhibited fair to good color fastness to
washing and light.

Acknowledgements
The authors wish to acknowledge the Support
Arts and Crafts International Centre of Thailand
(SACICT) for financial support throughout this
work.
References
1. EU-Thailand Small Projects Facility on
Industry Structure: Value Chain Analysis for
Thai Home Textiles: Silk Sub-sector Chiang
Mai, Thailand 2007.
2. Fredddi G, Mossotti R, Innocenti R.
Degumming of silk fabric with several
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4. Gulrajani ML, Gupta SV, Gupta A, Suri M.
Degumming of silk with different protease
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5. Gulrajani ML, Agarwal R, Grover A, Suri M.
Degumming of silk with lipase and protease.
Indian J. Fibre Textile Res. 2000, 25: 69-74.

Conclusion
By comparing Thai Bombyx mori silk fibers
degummed with papain enzyme and alkaline/soap
the former exhibited less tensile strength drop and
gave higher color depth after natural lac dyeing,
especially when degumming occurred at room
temperature condition. However, percent weight
loss of all degumming processes was within the
same range. SEM technique could be utilized to
verify the amount of remaining sericin after
degumming. As future work, dyeing with other

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6. Gulrajani ML, Agarwal R, Chand S.

Degumming of silk fungal protease. Indian J.
Fibre Textile Res 2000, 25: 138-142.
7. http://www.sehda.org/docs/papain.pdf
8. ASTM D 3822-01: 2001 Standard Test
Method for Tensile Properties of Single
Textile Fibers (Pennsylvania USA: the
American Society for Testing and Materials,
2001).
9. Schindler W, Finnimore E. Chemical
assessment of damage to fibers and fabrics.

In: Chemical testing of textiles (Edited by Q

Fan Q) Woodhead Textiles Series No. 40
2008, 193-194.
10. ISO 105-C01:1989 TextilesTests for color
fastness. Part C01: Color fastness to washing:
Test 1 (Geneva: International Organization for
Standardization, 1989).
11. AATCC Test Method 16 (2004) Colorfastness
to Light (North Carolina USA: American
Association of Textile Chemists and
Colorists, 2004).

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