Journal of Applied Biology & Biotechnology Vol. 11(4), pp.

44-49, Jul-Aug, 2023

Available online at http://www.jabonline.in
DOI: 10.7324/JABB.2023.40719

Bioprocessing of coir fibers using microbial consortium from termite

gut microbiota and Averrhoa bilimbi extracts

Revathy Rajan1* , Ajith Sudhakaran1 , Anita Ravindranath1 , Rajathy Sivalingam2, Ratheesh Kumar2
1
Department of Microbiology, Central Coir research institute, Coir Board, Kalavoor, Kerala, India.
2
School of Environmental Studies, Cochin University of Science and Technology, Kochi, Kerala, India.

ARTICLE INFO ABSTRACT

Article history:
Coir fiber, a tough natural lignocellulosic fiber, has widespread uses but are restricted due to its high tensile strength
Received on: December 16, 2022
and flexural rigidity. Bioprocessing, particularly softening and bleaching, is a common technique in the coir trade,
Accepted on: March 12, 2023
which currently uses chemical processes that are not environmentally friendly. The current approach targets coir
Available online: June 04, 2023
fiberprocessingwith Kozakonia oryzendophytica and Pseudomonas chengduensis isolated from the termite gutfor the
delignification followed by the treatment with Averrhoa bilimbi extract to obtain softened and bleached coir fibers.
Key words:
The physical and chemical assessment of the treated fibers, including variations in lignin content, flexural rigidity to
Softening,
Bleaching, assess softness, light fastness, Fourier-Transform InfraRed, and tensile strength, was carried out. The lignin presents
Tensile strength, in the treated coir fibers dropped significantly from 46% to 31.1%. The flexural rigidity was reduced from 1.12 to
Flexural rigidity, 0.53 which caused the upsurge in the degree of softness. The brightness index was increased from 9.45 to 13.49. The
Ecofriendly. scanning electron microscope images show that exterior protrusions on the fiber had been removed after treatment.
Thus, the present investigation discovers a trajectory for a biological method of coir fiber softening.

1. INTRODUCTION The color as well as the rigid property of the coir fiber are due to the
presence of a lignin network between fiber cells and are extremely
Coir is a versatile lignocellulosic natural fiber [1], with lignins and
impervious to biodegradation [5]. The high flexural rigidity due to
hemicelluloses forming the strengthening constituents of the fiber
the lignin content in coir fibers renders it unsuitable for a variety
cells. The tough fiber taken out from the husk of coconut is used to
of applications. Thus, lessening of flexural rigidity and toughness
make well-designed floor endowing furnishings in coir division set up
is vital for making coir fiber, anoutstanding material for numerous
in various coconut cultivating regions of India. These divisions supply
to the domestic and export marketplace [2]. The coir fiber used to make applications including geotextiles and Geomashes. Various chemical
mats, matting, runners, rugs, and carpets is whitened using a variety of approaches are employed for upgrading the quality of coir fiber,
reagents such as hypochlorites, peroxides, peracids, chlorites, sulfites, namely, smoothness, light fastness, and tensile strength for advanced
borohydrides, bisulfites, and others. Alkaline treatment is one of enactment of the coir fiber for commercial uses [6]. Biological cures
the most effective and low-cost procedures for softening coir fiber; are more practicable and ecofriendly which concurrently augments
however, it has been proven to be detrimental to the yarn’s strength. softening and bleaching. Although lignin deprivation by bacteria
Biosoftening uses specialized microorganisms with definite enzyme has been researched extensively in white-rot and brown-rot fungus,
specificity for topical cell wall elements to generate a biopolishing it is not wellunderstood in bacteria. Termites are one of the possible
effect on coir fibers. Unlike chemically softened fibers, the microbial sources of lignin-degrading machinery in nature. Few studies are only
treatment produced soft, brighter fibers with higher tensile strength reported regarding the digestive processes and the microbial associates
and elongation qualities, as well as superior durability [3]. Biosoftened residing the digestive gut of the fungus cultivating termites [7]. The
coir fiber is spinnable and may be combined with other natural fibers microflora in the termite gut digests lignocellulose into by-products
such as sisal, jute, and banana to create fabrics, textiles, and other such as molecular hydrogen and acetate and is consequently taken
products [4]. up by the termite or other microorganisms in the community. The
previous research has found that termite microflora removes 5–83% of
the lignin, along with most of the neutral polysaccharides and the best
*Corresponding Author: part of the acidic saccharides [8,9].
Revathy Rajan,
Department of Microbiology, Central Coir research institute, Coir Board, Averrhoa bilimbi L., commonly called bimbling plum, blimblin;
Kalavoor, Kerala, India. English: bilimbi, cucumber tree, tree sorrel; Filipino: Kamias; French:
E-mail: ammu55u @ gmail.com Blimblim, blinblin, and carambolier bilimbi [10], is a member of the

© 2023 Revathy Rajan, et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercial-ShareAlike
Unported License (http://creativecommons.org/licenses/by-nc-sa/3.0/).
 Rajan, et al.: Coir Fiber Bioprocessing with Termite Gut Microflora and A.bilimbi 2023;11(4):44-49 45

Oxalidaceae family [11], which is extensively grown in tropical areas. anthrone reagent (200 mg anthrone was dissolved in 100 mL ice-cold
The fruit juice contains more oxalic acid and hence used to eliminate 95% sulfuric acid and chilled for 2 h before use) was added and cooled
iron-rust colors from clothing and gives brassware, a lustrous and the absorbance was read at 630 nm [16,17].
sheen [12,13]. Biosoftened coir fibers find use in a variety of industries
since it is spinnable and effortlessly combined with other natural fibers 2.5. Surface Morphology
[4]. The current work seeks to bleach and soften coir fiber using a
The external modifications of the raw (untreated) and processed fibers
microbial consortium derived from termite gut microflora, followed
were checked with the help of FESEM (Carl Zeiss (USA), Model:
by A. bilimbi extracts, and then assess the hue and quality of the coir
Sigma with Gemini column, resolution 1.5 nm).
fiber. Research has been done on the processing of coir fibers using
ligninolytic fungi and bacteria, but no reports have been made yet on
exploiting the termite gut microflora in combination with the natural 2.6. Flexural Rigidity
fruit extracts to soften and brighten the coir fibers. This is the first The flexural rigidity of untreated and coir fibers was tested using a flexural
time Kosakonia oryzendophytica and Pseudomonas chengduensis, rigidity tester designed by CCRI. The raw and treated fibers, 25 samples
both isolated from termite guts, which have been used to improve coir each, were tied around a 2-inch diameter PVC pipe to make a ring.
fiber bleaching and softening. The present investigation leads to the A Flexural Rigidity Tester was employed to test the rings created by the
development of a bioformulation which could be used as an alternative fibers with and without stress (1 g). This information was used to calculate
to the present chemical methods of bleaching as well as softening the ring diameter and deformation during load suspension. Then, the
employed in the coir industry to soften the coir fibers. flexural rigidity was calculated by the following mathematical equation

2. MATERIALS AND METHODS Flexural Rigidity (gcm2) = 0.0047 • mg • (2πr)2• (cosθ/tanθ) [18].

2.1. Coir Fiber Treatment Where, mg =weight of the load in grams

Coir fiber (5 g) was weighed for each treatment, rinsed well in running r=radius of the ring in cm
water, and dehydrated extensively. Nutrient broth was prepared for
300 mL and inoculated with bacterial strains K. oryzendophytica d=deformation of lower endofthe ringincm
(Accession No. OM943747) and P. chengduensis (Accession No. θ=493d/2πr
OM943748) isolated fromthe gut of termite, Odontotermes obesus,
collected from the vicinity of Central Coir Research Institute, Kerala, 2.7. Tensile Strength
followed by the addition of coir fiber [14], which acts as a lignin
substrate for the microbes’ growth in the medium. The culture was then The tensile behavior of the fiber were tested employing the Universal
incubated at 37°C for up to 72 h. The treated fibers, after incubation, Testing Machine (UTM) Shimadzu AG-X/R with different parameters,
were completely washed, dried, and surface sterilized under UV light namely, strain rate = 10 mm/min, gripping length = 5 cm at atmospheric
before being treated with A. bilimbi. pressure.

2.2. Treatment with A. bilimbi Extracts 2.8. Lightfastness

In a rotary shaker, the microbially treated fibers were immersed in The Xeno test (Colorfastness to light, air-cooled xenon arc lamp), IS
A. bilimbi extracts and cultured for 72 h at room temperature. The standard AATCC 16 H-1998, was used to determine the lightfastness of
volume of extract used for the treatment was weighed for 150 mL to the treated fibers. It was done by exposing a fiber sample to a light source
ensure that all of the fibers were entirely soaked. The raw extract was for a certain amount of time and then comparing it to an unexposed
used in triplicate for the treatment. The material was rinsed in distilled sample. The Xenotest can also be used to determine fiber quality by
water, dried at room temperature, then analyzed, and then characterized. evaluating lightfastness ratings [16]. This is a permanent specification
for evaluating the light fastness and weather fastness of target samples
at a faster rate than is possible. A 1500-watt Xenon arc lamp is utilized
2.3. Estimation of Lignin
in the Xenotest to provide radiation; the cleared spectrum of this lamp
The Chesson method was executed to calculate the amount of lignin employed in the Xenotest is equivalent to sunlight. The fiber samples
in the samples [15]. After 30 min of drying in a hot air oven at 105°C, were exposed to light and dark for alternating intervals. This method
1 g of coir fiber was weighed. After shaking for 10 min with 5 mL of comes close to simulating real day and night circumstances. To assess
98% sulphuric acid, the sample was shifted to a 1000 mL Erlenmeyer color change, exposure was stopped when the contrast between the
flask with 450 mL distilled water. The mixture was boiled for 10 min exposed and unexposed portions of the specimen was equivalent to the
and filtered through a grade 1 glass filter. The residue was then washed greyscale grades. The tested and original fabrics were compared under
to neutrality and dried at 105°C and the final weight was measured.
a white light using the blue standard as a reference to determine color
change [Table 1] [19].
2.4. Estimation of Cellulose
One gram of the coir fiber sample was mixed with 3 mL acetic/nitric 2.9. Brightness Index
mixture (150 mL of 80% acetic acid + 15 mL concentrated nitric acid)
The brightness index of the fibers was verified as per the TAPPI-452
utilizing a vortex mixture and kept in a waterbath for 30 min at 100°C.
method with the help of premier color scan spectrophotometer
After cooling, the mixture was centrifuged for 15 to 20 min. After
(SS 5100A).
the supernatant was drained, the residue was carefully washed with
distilled water. The washed residue was mixed with 10 mL of 67%
sulfuric acid and allowed to stand for 1 h and later diluted 1 mL of 2.10. Fourier-Transform InfraRed (FTIR) Spectroscopy
thesolution to 100 mL. To 1 ml of the above-diluted solution, 10 mL of Comparative analysis and identification of surface structural groups
46 Rajan, et al.: Journal of Applied Biology & Biotechnology 2023;11(4):44-49

Table 1: The light fastness grades [15].

Grade Degree of fading Light fastness type
8 No fading Outstanding
7 Very slight fading Excellent
6 Slight fading Very good
5 Moderate fading Good
4 Appreciable fading Moderate
3 Significant fading Fair
2 Extensive fading Poor
1 Very extensive fading Very poor

Figure 1: Averrhoa bilimbi fruit (extract) used to obtain softened and

bleached coir fibers.
b
of control and treated coir fibers wereperformed by Thermo Nicolet, Figure 2: Estimated cellulose and lignin contents in coir fiber after
Avatar 370 FTIR spectrometer. With a resolution of 4 cm−1 and 32 treatment using Pseudomonas chengduensis + Averrhoa bilimbi, Kozakonia
scans per sample, FTIR spectrum of the raw fiber and microbial oryzendophytica + Averrhoa bilimbi, and Consortium + Averrhoa bilimbi
treated fibers was taken and the absorbance spectra were recorded at at different hours. (a) Cellulose and (b) Lignin percentages were obtained
wavenumbers from 500 to 4000 cm−1. after every treatment tested at different hours. Six replications (Mean ± SE)
were maintained for both estimations. Vertical bars under different small
2.11. Statistical Analysis letters were statistically significant, and the same letters indicate statistically
Two-way ANOVA was performed using IBM SPSS v 20 (IBM insignificant (Tukey test, LSD; P < 0.05) among the treatments (Pseudomonas
Corporation, Armonk, NY) to analyze the significant difference (Tukey chengduensis, Kozakonia oryzendophytica, and Consortium). Different
test, LSD; P < 0.05) in cellulose and lignin contents retrieved from coir capital letters on the top of the vertical bar indicate that every treatment tested
fiber after different treatments (P. chengduensis, K. oryzendophytica at different hours (24, 48, and 72) was statistically significant (Tukey test,
and Consortium) tested at different hours (24, 48, and 72). LSD; P < 0.05), and the same capital letters indicate statistically insignificant.

3. RESULTS AND DISCUSSION of extracellular enzymes and secondary metabolites by the bacterial
Changes in the degree of softness and vividness of the coir fibers are species makes them, the major lignocelluloses degraders in
approved by the treatment. The hard lignin network that connects soils [23,24]. Researchers reported that the wood-feeding termites
fiber cells inhibits the fiber from becoming flexible. Since lignin is found in tropical savanna and forests are able to digest lignocellulosic
the component responsible for color and stiffness, fiber softening with substrates efficiently and, hence, likely have a lignin-preconditioning
negligible impact on fiber strength can be attained in conjunction machinery that allows them to cope such proficient deprivation of
with the biobleaching treatment [20]. The surface of coir is plentiful timbered floras [25]. Based on the reports by Rajan et al., the bilimbi
fruit extract has the potential to lower the stiffness of coir fibers
with amorphous lignin, and the inside structure is sculpted by cell
[Figure 1] [15].
walls, which are abundant in crystalline cellulose, according to the
previous reports [16]. Researches have reported the development of
plasma treatment for coir fibers that involved etching the fibers [21]. 3.1. Lignin and Cellulose
In nature, lignin breakdown occurs as a result of collaboration between Estimated cellulose and lignin contents in coir fiber after treatment using
fungi and bacteria in natural microbial consortia [22]. The production P. chengduensis + A. bilimbi (Pc+Ab), K. oryzendophytica + A. bilimbi
 Rajan, et al.: Coir Fiber Bioprocessing with Termite Gut Microflora and A.bilimbi 2023;11(4):44-49 47

(Ko+Ab), and Consortium + A. bilimbi (C+Ab) were compared among the oxalate present in the bilimbi extracts may be accountable for the
the treatments and the performance of individual treatments at different partial delignification of coir fibers. The outcome disclosed the ability
hours. After being treated with the microbial consortium and then of the method to diminish the inflexible arrangement of coir fibers.
bilimbi, it was discovered that coir fiber had less lignin on the surface,
resulting in changes in softness. The lignin in coir fiber (control) was 3.2. Surface Morphology
lowered on the combined treatment at various intervals of incubation,
External micropore protrusions on coir fibers may provide defianceand
as shown in Figure 2. The Pc+Ab performed significantly at 24 h of
toughen the coir fiber surface. It is evident from the results that the
treatment, and the percentage of cellulose content estimated from
apparent deposit of the coir fiber has been wiped on treatment and
coir fiber was significantly high compared with Ko+Ab and C+Ab
the internal packs of micro fibrils are observable. Thus, the external
treatments [Figure 2a]. The treatments were extended to 48 and 72 h and
micropore protrusions have been cleared through the process resulting
did not significantly differ; loss in cellulose percentage is significantly
the fiber getting smoother. The SEM pictures and descriptions disclose
low [Figure 2a]. On the other hand, the treatments of coir fiber with
the exclusion of the external cover which is abundant with cellulosic
C+Ab showed a significant decrease in the percentage of lignin when
groups and amorphous lignin that uncover the core arrangement of
compared to individual treatments using Pc+Ab and Ko+Ab [Figure 2b].
packs of cellulosic microfibrils [Figure 3].
The cellulose composition was found to be reduced insignificantly
which proves that the method effectively softens and brightens 3.3. Flexural Rigidity
the fibers with negligible loss of the cellulose content of the fibers
When the fibers were treated with the microbial consortium followed by
[Figure 2a]. Meanwhile, the lignin concentration was observed to be
A. bilimbi treatment at 72 h of incubation, the flexural rigidity decreased
reduced by up to 32% after treatment with microbial consortia and
significantly from 1.12 (for untreated fiber) to the lowest of 0.53
bilimbi extracts after 72 h of incubation. The reciprocal action of the
[Table 2]. The outcomes show that treatment with the microbial consortia
lignolytic enzymes produced by the microbial consortia as well as
modifies the apparent structures of coir fibers without affecting the core
structure or strength. Thus, the treatment is a favorable, economical, and
biological technique for attaining softened coir fibers.

3.4. Tensile Strength, Brightness Index, and Light Fastness

The breaking stress imparted on raw fiber and treated fibers was found
to be lowered from 245.35 to 230.999, indicating that the treatment
had relatively minor effects on fiber strength [Table 2]. The brightness
a b index of the coir fibers increased from 9.45 to 13.49 after treatment with
the microbial consortia and A. bilimbi [Table 3]. Due to lignin build-up
on the fiber surface, the fiber darkens, necessitating bleaching, which
makes the fiber brittle. The strength of chemically softened fibers is
dramatically diminished, which has a negative impact on spinning
characteristics and yarn strength. The light fastness of the fibers
improved from Grade 2/3 to Grade 3/4 after the combination treatment
[Table 3]. Studies on biobleaching of jute (Indian Jute Industries
c d
Research Association, Annual report, 1984–1985) found that in situ
Figure 3: Scanning electron micrographs of raw and treated fibers. (a) Raw growth of selected lignolytic white-rot fungus Polyporus sanguineus
fibers. (b) Pseudomonas chengduensis + Averrhoa bilimbi extract treated on raw jute and grey fabrics improved the light fastness of the samples
fiber, (c) Kozakonia oryzendophytica + Averrhoa bilimbi extract treated (Indian Jute Industries Research Association, Annual report, 1984–
fiber, and (d) Microbial consortium + Averrhoa bilimbi extract treated fiber. 1985) [4]. Biosoftening softens, thins, and bleaches the fiber while
(Magnification at ×500). avoiding the use of caustic chemicals, resulting in less pollution [3].

Table 2: Tensile strength and flexural rigidity of raw and treated coir fibers after 24, 48, and 72 h of incubation.
Sample Time of incubation (hours) Tensile strength Flexural rigidity
Breaking stress (N/mm2) Break strain (%) Flexural rigidity (Gf)
Raw coir fiber 245.351 25.024 1.12
Pseudomonas chengduensis+Averrhoa bilimbi 24 241.346 25.425 0.89
48 227.943 24.764 0.81
72 213.048 26.946 0.78
Kozakonia oryzendophytica+Averrhoa bilimbi 24 239.907 25.678 0.91
48 224.053 26.543 0.87
72 217.092 26.386 0.77
Microbial Consortium+Averrhoa bilimbi 24 245.610 25.703 0.65
48 212.145 26.079 0.59
72 230.999 24.243 0.53
48 Rajan, et al.: Journal of Applied Biology & Biotechnology 2023;11(4):44-49

Table 3: Light fastness and brightness index of raw and treated fibers after the gut micro flora of termites to delignify and soften the coir fibers and
72 h of incubation. the bleaching property of A. bilimbi to impart brightness to the fibers.
Sample Light Brightness The combined treatments using the consortium and fruit extracts have
fastness index functioned well in transforming the surface topography, drop in the
type lignin proportion, flexural rigidity, and augmentation of the brightness
Raw fiber 2/3 9.45 index. The technique commends that the cure does not affect the inner
assembly of coir fiber but modifies the external alignments of lignin
Treated fiber at 72 h incubation
thereby refining the coir fiber eminence without losing strength. Thus,
Pseudomonas chengduensis+Averrhoa bilimbi 3 10.25 the treatment will slash its trajectory to a bio method, an alternative for
Kozakonia oryzendophytica+Averrhoa bilimbi 3 10.49 the prevailing chemical methods, and thus benefits the future coir trade
Consortium+Averrhoa bilimbi 3/4 13.49 to be more environmentally friendly.

5. AUTHOR’S CONTRIBUTIONS

All the authors have made extensive scholarly contributions to mount

this manuscript in the following areas: Conceptualization: Revathy
Rajan, Ajith Sudhakaran, Anita Ravindranath, Rajathy Sivalingam
and Ratheesh Kumar; data acquisition/analysis/interpretation/
drafting manuscript: Revathy Rajan and Ajith Sudhakaran; Critical
Revision of the manuscript: Revathy Rajan, Ajith Sudhakaran, Anita
Ravindranath, Rajathy Sivalingam and Ratheesh Kumar; supervision
and final approval: Anita Ravindranath, Rajathy Sivalingam and
Ratheesh Kumar.

6. FUNDING
There is no funding to report.

7. CONFLICTS OF INTEREST

Figure 4: FTIR spectra of raw fiber and coir fiber treated with microbial The authors announce that they have no conflicts of interest.
consortium together with Averrhoa bilimbi extract.
8. ETHICAL APPROVALS
3.5. FTIR Spectroscopy The research work does not include experimentations on animals or
Primarily, the peak ranging from 3000 cm−1 to 3700 cm−1 is related human subjects.
to the stretching of hydroxyl groups [26]. Given the spectrum of
the untreated coir fiber as a citation to check, the efficiency of the 9. DATA AVAILABILITY STATEMENT
microbial + bilimbi cures performed on the fiber, for the spectrum of
treated fibers, the deepened bands such as a 1024-cm−1 bond bound to All datasets were generated and analyzed in the present study.
a C–H stretching vibration of the cellulose backbone and 2908 bonded
to an ester bond, and the band 3328 cm−1 associated to the existence 10. PUBLISHER’S NOTE
of hydroxyl groups (–OH) of cellulose. Meanwhile, the absorption This journal remains neutral with regard to jurisdictional claims in
band corresponding to the C = O stretching of carboxyl and acetyl published institutional affiliation.
groups in the hemicellulose generated the untreated coir fiber peak
band at 1738 cm−1. However, it shrank when the fibers were treated
11. ACKNOWLEDGMENTS
with the consortium + A. bilimbi which relates to the elimination
of hemicellulose, and also the band near 1228 cm−1 representing The University Grants Commission, New Delhi, India, and the
the presence of CH stretching of acetyl groups of lignin exhibited a Kerala State Council for Science Technology and Environment,
direreduction in the intensity in the treated fibers [27]. The region a self-governing agency of the Government of Kerala, have both
between 1252 and 1439 cm−1 is associated with the lignin content generously supported the authors. The authors also acknowledge
category of the C = H group [28,29]. Fascinatingly, the reduced peak Shri. D. Kuppuramu, Chairman, Coir Board, for expanding CCRI’s
intensity at 1368 cm−1 specifies the elimination of surface lignin in laboratory capabilities. The School of Environmental Studies at Cochin
the treated fibers. These alterations show the exclusion of lignin and University of Science and Technology, Cochin, India, also provided
hemicellulose from the coir fiber surface and, hence, attest that the assistance and facilities, in which the authors gratefully acknowledge.
treatments were done effectively to alter the physiognomies of the We sincerely appreciate the time and assistance given to mount the
fiber surface [Figure 4] [22]. paper by Shri. Sivaprasath Prabhu, Chinese Academy of Agricultural
Sciences Institute of Plant Protection, Beijing, China.
4. CONCLUSION
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