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Papain enzymatic degumming of thai Bombyx mori silk fibers

Article · January 2009

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 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: [email protected]

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.

Background Papain is a proteolytic enzyme produced from

Thai Bombyx mori silk, produced by cultivated the latex of the green papaya fruits [7]. It has high
mulberry silkworm, is greatly appreciated as one of protein hydrolyzing capacity. Major applications of
the most luxurious textile commodities. Known for papain enzyme include food industries, primarily
its unique characteristics such as luster and for chill proofing of beer, tenderizing of meat and
softness, the production of Thai Bombyx mori silk freeing of food proteins. Other applications are in
consists of silkworm raising, cocoon production, tanning of leather and hides, degumming of silk,
silk reeling, degumming, dyeing, weaving, and cheese manufacture, pharmaceuticals, aroma and
finishing products [1]. Degumming is the process perfume industries and effluent treatment.
where sericin, i.e., the silk gum gluing the fibroin The aim of the present research is to investigate
filaments, is totally removed in order to obtain a the influence of papain enzymatic degumming of
typical shiny, soft handle and other desirable Thai Bombyx mori silk fibers on fiber damage and
properties [2]. Due to its highly oriented and residual sericin by percent weight loss calculation,
crystalline structure, fibroin is not water soluble, tensile strength and scanning electron microscopy
whereas sericin is easily dissolved in a boiling (SEM) measurement and color staining testing. The
aqueous solution with the presence of soap, alkali, traditional alkali/soap degumming method was also
organic acid or synthetic detergents. Nowadays, performed for comparison. In additions, the
batch degumming of silk is mostly carried out in an resultant degummed silk fibers were dyed with a
alkaline bath with pH about 10 containing 10-20 natural lac dye in order to evaluate their dyeability.
g/L soap at 92-98 °C for 2-4 h [2-3]. During highly
hot alkaline silk degumming, hydrolytic Materials and Methods
degradation of fibers is accompanied, leading to Materials
undesirable both aesthetic and physical properties The Bombyx mori silk fiber used in the
such as dull appearance, surface fibrillation and experiment was sourced from the northeast part of
tensile strength loss. Thus, synthetic detergents Thailand. Papain enzyme was derived from
may be used to replace soaps in case of reduced papyrus latex of Thai’s green papaya fruits with
treatment time and less fiber damage are needed. 1.5-3 months of age. The collected latex was
Furthermore, enzymatic treatment of silk is successively dried in the open air and in the oven at
currently gained large attention as an alternative for 60 °C and then ground into a yellowish powder.
degumming process owing to its ecologically safe Lac dye was obtained from the northeast of
and required milder conditions [4-6]. Proteases and Thailand. Alum (AlK(SO4)2.12H2O) was purchased
lipases are normally used in combination for from Asia Pacific Specialty Chemicals Ltd.,
degumming and removing others impurities such as Thailand. All other chemical reagents were of
waxes, fats, mineral salts and pigments [2-3]. 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

Silk degumming with alkali/soap dyed with a lac dye solution in a liquor ratio of
Silk samples were treated with an alkaline bath 1:30 at 90-95 °C for 45 min. Dyed silks were then
containing an aqueous solution of 10 g/L soap and rinsed with water and air-dried.
2 g/L sodium carbonate in a liquor ratio of 1:30 at Color measurement and testing
90-95 °C for 45 min. Degummed silks were then The color strength (K/S values) and CIELAB
washed with hot and cold distilled water and finally (L*a*b*) values of dyed silk fibers were measured
air dried. on the Gretag-Macbeth Color i5 spectrophotometer
Silk degumming with papain enzyme with illuminant D65 at 10 degree observer. Color
In our preliminary experiment, it was found that fastness tests to washing and light were determined
both papain enzyme concentration and condition according to ISO 105-C01 (1989) and AATCC
(temperature and time) played crucial roles in order Test Method 16 (2004), using DaeLim Starlet II IR
to remove sericin from the silk fibers. In addition, lab dyeing machine and Suntest CPS testing
the optimum concentrations of papain enzyme were equipment, respectively [10-11].
at 5% and 10% owf were required for degumming
at 65-70 °C for 1 h and at room temperature for 24 Results and Discussion
h, respectively. Hence, in this work, silk samples Table 1 summarizes the results on efficiency of
were degummed according to the above deduction. alkaline/soap and papain enzymatic degumming
Degummed silks were then washed with hot and processes of silk fibers in terms of percent weight
cold distilled water and finally air dried. loss, breaking load and color staining. Although
Determination of the weight loss there was a similar reduction in percent weight loss
The percent weight loss of the silk fibers after (i.e., approximately 20-22%) of Bombyx mori silk
degumming process was calculated from Eqn 1: fibers degummed with alkali/soap and papain
enzyme, a distinct difference in breaking load was
 W 0 -Wt  clearly observed. Degumming silk fibers with 5%
% Weight loss =   ×100 (1) owf papain enzyme at high temperature (65-70 °C
 W0  for 1 h) yielded a maximum decrease of breaking
load, i.e., about 46% of the raw silk. On the other
where W0 and Wt are the weight of the silk fiber hand, the breaking loads of silk fibers degummed
samples before and after degumming, respectively. with 10% owf papain enzyme at room temperature
Determination of breaking load for 24 h and with alkali/soap dropped to 28.4% and
Breaking loads of silk fibers were performed 40.2%, respectively. This was probably due to an
according to ASTM D 3822-01 Standard Test intense enzyme activity at a higher temperature
Method for Tensile Properties of Single Textile condition compared to at room temperature.
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 Table 1 Percent weight loss, breaking load and color
staining results of Bombyx mori silk fibers.
LR5K, Intro Enterprise Co., Ltd.).
Degumming Weight Breaking Staining
Scanning electronic microscopy (SEM) analysis Methods loss (%) load (N) Test
Morphological characterization of silk fibers
was performed by means of scanning electron Raw silk - 8.0540 Dark red
microscopy (Model 5410, JEOL). Samples were Alkaline
observed at 15 kV acceleration voltage after gold 21.94 4.8190 Pale pink
/Soap1
sputtering. Papain
20.60 4.3475 Pale pink
Color staining test 5 %owf 2
Ambidirect Red 3BL 165% (C.I. Direct Red Papain
20.80 5.7632 Pale pink
80), supplied by Thai Ambica Chemicals Co., Ltd 10 %owf 3
was used to evaluate the remaining of sericine after 1
90-95 °C, 45 min
silk degumming. The degummed silk samples were 2
65-70 °C, 1 h
immersed in an aqueous solution of 1 g/L 3
room temperature, 24 h
Ambidirect Red 3BL 165% with a liquor ratio of
1:200 for 2 min at the boil [9].
Lac dyeing For the direct dye staining test, raw silk was
Crushed stick lac was first extracted by water stained dark red color as a result of a large quantity
(weight ratio = 1:10) at the boil for 1 h and then of sericin present in the fibers. Both papain and
filtered to give a red solution of lac dye. The lac alkali/soap degummed silk appeared pale pink,
dye solution showed a maximum absorption peak indicating that there were a small amount of sericin
(λmax) at approximately 485 nm. The degummed remaining. Moreover, dull and stiff handle of raw
silk fibers were pre-mordanted with 10% owf alum silk fibers disappeared and the degummed silk
solution at room temperature for 30 min and then became more shiny and softer.

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

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

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 Dyed CIELAB Color fastness
K/S
methods samples L* a* b* Washing Light

Alkaline
6.17 44.65 38.78 1.39 3 3
/Soap

Papain
8.72 37.28 35.29 0.64 2-3 2-3
5 %owf

Papain
9.78 35.86 34.23 1.34 2 2-3
10 %owf

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)

SEM observation reveals that the raw Bombyx natural dyes could be also undertaken as another
mori silk fibers were covered with a relatively large assessment of the efficiency of papain enzymatic
amount of sericin on fibroin as illustrated in Figure degumming.
1. Degumming of the fibers with alkali/soap and
papain enzyme at 65-70 °C or at room temperature Acknowledgements
could almost completely remove sericin. However, The authors wish to acknowledge the Support
fibrillation of some fibroin threads occurred during Arts and Crafts International Centre of Thailand
the degumming processes. (SACICT) for financial support throughout this
In view of natural lac dyeing, papain enzymatic work.
degummed silk fibers had higher color strength
(K/S values) than those degummed with alkali/soap References
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 Journal of Microscopy Society of Thailand 2009, 23(1): 142-146

6. Gulrajani ML, Agarwal R, Chand S. In: Chemical testing of textiles (Edited by Q

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Method for Tensile Properties of Single Test 1 (Geneva: International Organization for
Textile Fibers (Pennsylvania USA: the Standardization, 1989).
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