Cleaner Engineering and Technology 5 (2021) 100304

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Cleaner Engineering and Technology

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Eco-friendly degumming of natural fibers for textile applications: A

comprehensive review
Mira chares Subash , Perumalsamy Muthiah *
Department of Chemical Engineering, National Institute of Technology, Tiruchirappalli, India

A R T I C L E I N F O A B S T R A C T

Keywords: Global fiber production contributes 107 million MT in 2018 and is expected to grow 145 million MT by 2030.
Natural fiber Fiber production can be plant-based, animal-based, man-made, and synthetic fibers. Plant-based fibers include
Enzymes jute, ramie, and hemp, have a market share of 5.7% in textile industries. The emerging interest in plant-based
Degumming
fibers, agricultural residues could be addressed by adopting unexplored plant fibers. Textile processing of nat­
Textile industries
ural fiber necessitates the removal of the hemicellulosic substance. The major concern of this investigation is the
degumming of natural fibers by an eco-friendly method. The ideology is focused on producing hemicellulose
degrading enzymes through solid substrate fermentation and utilization of hemicellulose degrading enzymes in
natural fiber degumming. Degumming of fiber can be performed by hemicellulose degrading enzymes such as
Pectinase, Xylanase, and Laccases. The neglection of the non-cellulosic layer corresponds to an increase in tensile
strength, crystallinity, fineness, removal of non-cellulosic substances. The degummed fiber has remarkable ap­
plications in fabrication, cutlery application, and textile industries. Our investigation enclosed consolidated
reports on the agricultural residues for hemicellulose degrading enzyme production and application of enzymes
for natural fiber degumming proclaimed in recent years. The practical difficulties in adopting degummed fibers
in textile industries have also been discussed.

1. Introduction consolidates the degumming of plant-based natural fibers for textile

Fibers are thread-like structures with continuous or discrete phases
of filaments. The predominant advantage of natural fiber is biodegrad­
ability, lightweight, and less density. Natural fibers account for innu­
merable benefits of sustainability and eco-friendliness. The concern
about natural fibers research has increased recently due to the need for
an alternative towards synthetic fiber usage. Some cost-effective rein­
forcement materials from the natural fibers pre-owned as an alternative
to synthetic fibers are flax, jute, ramie, hemp, banana, etc. (Chabba
et al., 2005). The natural fiber application has been inclined tremen­
dously since the last decade. Jain et al. (2016) reported that the dislo­
cation of famous designers and renowned fashion brands was searching
for eco-friendly fibers to make sustainable products for clothing and
other apparel. Natural fibers account for cost-effectiveness, manageable
processing, and renewable resources. The natural fibers can be classified
as plant-based, animal-based, and mineral-based. The plant-based fibers
can be further categorized as bast fibers, leaf fibers, wood fibers, fruit
fibers, seed fibers, straw fibers, and grass fibers. Our research paper
applications.
1.1. Plant-based fibers
Seed fibers: Among the natural fibers, Cotton fiber is a highly
explored natural fiber. The source of cotton fiber is plant seeds, pre­
dominantly known as seed fibers. Cotton fiber belongs to the genus
Gossypium. Cotton fibers are enriched with cellulose. The 90% compo­
sition of Cotton fiber possesses cellulose. The characteristic features of
cotton fiber are absorbent, lightweight, and soft. The major applications
of cotton fiber are textiles, woven fabrics, fishing nets, and so on. Other
seed-based fibers are Pine-cone, Kapok, Loofah, and rapeseed (Pinheiro
et al., 2020).
Bast fibers: The members of bast fibers are Flax, Jute, Ramie, Kenaf,
Hemp, Mesta, and Roselle. Hemp fibers are identified to be one of the
strongest members of bast fibers. Hemp fibers are derived from the
species Cannabis. The composition of cellulose in hemp fiber is about
70%. Ramie fibers are powerful bast fibers with long durability and

* Corresponding author. National Institute of Technology, Tiruchirapalli, India.

E-mail address: mpsamy@nitt.edu (P. Muthiah).

https://doi.org/10.1016/j.clet.2021.100304
Received 25 March 2021; Received in revised form 16 October 2021; Accepted 17 October 2021
Available online 19 October 2021
2666-7908/© 2021 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
M. Subash and P. Muthiah Cleaner Engineering and Technology 5 (2021) 100304

higher cellulose content. Ramie fibers are also known as china-grass,
grass linen, grasscloth, and china linen. The durability and strength of
ramie fibers promote its application in industrial sewing thread, pack­
aging materials, fishing nets, and filter cloths.
Similarly, Jute fibers, commonly known as lignocellulosic bast fibers,
belong to the genus Corchorus. Kenaf fibers are derived from the bast of
Hibiscus comes under the family Malvaceae. The features of kenaf fibers
are flexural strength, lightweight, ignition, biodegradability, and eco-
friendliness (Jain et al., 2016).
Grass fibers: The grass fibers are Bamboo, Bagasse, Corn, Sabai, and
Canary. The bamboo fibers are derived from Bamboo plants that belong
to the family Bambusoideae. The bamboo fiber has enormous micro-gaps,
which are responsible for higher absorption quality. The features of
bamboo fibers are antimicrobial, hygroscopic, and resistant against UV
light.
Fruit fibers: Coir fiber and Oil-Palm fiber comes under fruit fibers.
Coconut or coir fibers are obtained from the husk of the coconut. The
coir fiber belongs to the family Cocus nucifera. The properties of coir
fibers are durable and cost-effective. Oil-palm fibers are also known as
lignocellulosic fibers, comes under the family Arecaceae. The source of
oil-palm fibers is fruit mesocarp and empty fruit bunch of oil-palm tree.
Leaf fibers: The leaf fibers are Sisal, Banana, Abaaca, Pineapple,
Henequen, and Agave. Banana fiber (lignocellulosic fiber) is generated
by the pseudostem, which has significant mechanical properties (Dra­
hansky et al., 2016). Banana pseudostem seems to be clustered, cylin­
drical solicitation of leaf stalk bases. The banana pseudostem can be
implemented for making marine rope, coffee, tea bags, filter cloths, and
light-density woven fabrics. (2010) mentioned the consideration to­
wards banana pseudostem in textile, paper, and composite material for
structural reinforcement. The pseudostem exhibits antimicrobial prop­
erties (Bello et al., 2018). Sisal fibers are the cellulosic fibers derived
from the Agave sisalana. The sisal fibers are perennial succulents with
major applications in rope-making, paper, cloth, geotextiles, dartboards,
footwear, hats, and bags. Sisal fiber belongs to the family Asparagaceae.
Wood fibers: Wood fibers can be classified as softwood and hard­
wood fibers. The major components of wood fibers are lignin, cellulose,
hemicellulose, and extractives. Both kinds of wood fibers play a major
role in paper manufacturing. The properties of natural fibers such as
banana, coir, cotton, flax, hemp, jute, kenaf, palm, pineapple, ramie,
sisal, bamboo, and dates are listed in the following Table 1 (Paluvai
et al., 2014) (Sreenivas et al., 2020).
1.2. Need for fiber degumming
Industrialization of natural fibers necessitates removing non-
cellulosic content of the threads, which degumming fibers can accom­
plish. Natural fibers, namely jute, ramie, kenaf, and hemp, have signif­
icantly experimented for fiber degumming application and production
of value-added products (Chiliveri et al., 2016). Banana fibers are found
to be less explored natural fibers in the degumming application.
Degumming eliminates the viscous or sticky consistency from the
cellulosic part of plant fibers to increase textile manufacturing in­
dustries. The techniques of degumming methods are acid, alkali, steam
explosion, ultrasonic, microwave, bacteria, fungus, and enzyme (Shen
et al., 2015). The microbial degumming has declined the chemical and
energy consumption of natural fiber.
1.3. Microbial degumming
The enzymatic degumming process is an eco-friendly method as it
reduces fiber damage without altering the properties of cellulosic fibers
(Patidar et al., 2018). The pectinase and xylanase were identified to be
adequate enzymes for fiber extraction and degumming process (Tibolla
et al., 2014) (Jacob et al., 2008). In microbial degumming, fibers will be
autoclaved for 20 min. After sterilization, degumming temperature,
degumming time, and enzymatic dosage for the degumming of natural
fibers will be optimized. The microbial pectinase is noteworthy in fiber
processing as plant cambium composes 40% pectin in dry weight
(Bruhlmann et al., 1994). The hemicellulose and lignin removal from the
fiber surface results in single fiber formation, which will be further
processed for textile applications(Angelini et al., 2015). Kohli et al.
(2019) confirmed the eco-friendliness and energy conservation by
adopting the pectinolytic enzymes in plant fibers degumming. (2013)
suggested the highlights of utilizing the nanoparticle-based pectin
degrading enzymes to reduce sugars and further application in the
degumming of ramie fiber.
2. Pectinase
Pectinases can be of different forms based on their activity on the
structure of polygalacturonate. Enzymatic activity differs concerning
the degree of methylation. The enzymatic and specific activity of pec­
tinase relies on the methylation degree. The chrysanthemum family was
identified to be the first structure of pectinase. β-helix, distinct β-sheets
from β strand turns are the structural topology of the pectinase enzyme
(Henrissat et al., 1995). The active sites are His223 and Asp201, which
act as a donator of proton and nucleophile, respectively. His223 pro­
vokes proton transfer, glycosides bond breakage, and generation of the
covalent bond. Simultaneously, the opponent residue restores the
enzyme active site by locating the H2O molecule to rescue the second
product (Palanivelu, 2006). Fig. 1 illustrates the 3-dimensional image of
the pectinase enzyme.
Some of the prevailing microorganisms involved in pectinase pro­
duction are Pseudomonas, Xanthomonas, Erwinia, Actinomycetes, and
Streptomycetes and fungal sources such as Aspergillus niger, Aspergillus
versicolor, Aspergillus flavus, Fusarium oxysporum, Rhizopus stolonifer,
Mucor racemous, Mucor hiemalis, Penicillium jenseni, Penicillium citrinum
and Trichoderm aviride (Priya and Sashi, 2014). The convenience of
microbes results in the decline of fiber damage, energy consumption,
and non-toxic effluents. Banik et al. (2008) put forward the exploitation

Table 1
Properties of Natural fibers.
Fiber type Density (Kg/m3) Cellulose (wt %) Lignin (wt %) Tensile strength (Mpa) Young modulus (Gpa) Elongation (%) Moisture content (wt %)

Banana 1,350 63–64 15 529–914 27–32 1–3 13

Coir 1,150 32–45 40–55 140–593 4–6 25–30 8
Cotton 1,600 85–90 0.4–1 287–597 5.5–12.6 7–8 7.85–8.5
Flax 1,500 71 2.2 500–1500 27.6 2.7–3.2 8–12
Hemp 1,480 70–74 3.7–5.7 550–900 30–70 1.6–1.8 6.2–12
Jute 1,460 61.1–71.5 12–13 393–773 26.5 1.5–1.8 12.5–13.7
Kenaf 1,400 45–57 8–13 930 53
Palm 0.7–1.5 42.7–65 13.2–25 50–400 0.57–9 2.5–18
Pineapple 1,440 70–82 5–12.7 640 2.4 11.8
Ramie 1,500 68.6–76.2 0.6–0.7 400–938 61.4–128 3.6–3.8 7.5–17
Sisal 1,450 66–78 10–14 511–680 9.4–22 2.0–4 10–22
Bamboo 910 503 35.91 1.4
Date 990 309 11.32 2.73

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M. Subash and P. Muthiah Cleaner Engineering and Technology 5 (2021) 100304

profitability compared to other methods. Commercial alkaline pectate

lyase is more capable of flax retting with high tenacity and low struc­
tural damage (Jain et al., 2016). Kaur et al. (2016) disclosed Bacillus
pumilus BK2 as the valuable cotton scouring source. Alkaline pectinase
has great scrutiny in the textile industry to replace water loss, energy
demand, and toxic chemicals. They play a major role in degumming bast
fibers, ramie fibers, flax fibers, and other eco-friendly fibers. The syn­
thesis of pectinases from various species under different operating
conditions and their advantages are listed in Table 3.

3. Xylanase

Xylan, a polysaccharide, constitutes β-xylopyranose residues as spi­

Fig. 1. The 3-D image of Pectinase enzyme.
of pectinolytic microbes in the piling of jute fibers and their ability to
enhance yarn quality. The propensity of pectinases in the breakdown of
complex molecules, retting, and degumming of fiber crops are remark­
able (Chiliveri et al., 2016).
2.1. Production of pectinase by fermentation
In contrast to submerged fermentation, the supremacy of low
contamination, high yield, utilization of agricultural residue as sub­
strates, simplicity of processing, and lower energy requirements were
the convenience of solid-state fermentation (Selvakumar et al., 1998).
The solid substrate fermentation process is more than 5.87 times ad­
vantageous than other processes (Kapilan and Arasaratnam, 2011).
Investigation on the substrates suggests sugar cane bagasse, wastes of
tea, cassava, apple, oil mill, banana, corn, wheat (straw, bran, husk,
flour), coconut coir pith, rice (straw, bran), sawdust, the pulp of sor­
ghum and sugar beet, cake of rapeseed, coconut oil, etc. for the
fermentation. The Pectinase production using various species and sub­
strate are listed in Table 2.
2.2. Application of pectinases in degumming of fiber
Textile application of natural fibers prerequisite the degumming
process, which detaches the non-cellulosic material. The enzymatic
degumming overwhelms the pitfall of alternate methods (Baracat et al.,
1991). Kashyap et al. (2001) reported that Bacillus enzyme treatment
was used in the retting of Mistumatabast fibers, yielding high
nal with glycosidic linkages. Xylan and pectic substances are inter-
linked by xylan-glucan-protein complexes (Selvendran, 1985). The
xylan was found to be aggregated with hemicellulose via
hydrogen-bonding interactions. The xylanase enzyme system comprises
the repertoire of hydrolytic enzymes which act cooperatively for the
conversion of xylan into constituent sugars such as acetyl xylan esterase,
β-xylosidase, α-L-arabinofuranosidase, α-glucuronidase, β-1,4-endox­
ylanase, and phenolic acid (ferulic and p-coumaric acid) esterase. Pro­
duction of xylanase can be by bacteria, fungi, yeast, and marine algae.
The inherent sources of xylanase were identified to be Bacillus among all
the bacterial species. The other species of Bacillus were also reported in
terms of considerable xylanolytic activity (Gupta et al., 2015). Tricho­
derma reesei LM-UC4 E fungus was mixed with Aspergillus niger ATCC
10864 and A. phoenicis QM 329 to produce xylanase using Solid-state
fermentation (Rabello et al., 2014). The three-dimensional image of
the xylanase enzyme is illustrated in Fig. 2.
3.1. Production of xylanases by fermentation
Solid substrate fermentation is becoming an attractive process in
xylanase enzyme production because of its higher productivity and
lower synthesis cost. Using lignocellulosic materials in place of com­
mercial xylan in the fermentation can deduct the synthesis cost of
xylanase (Senthilkumar et al., 2005). The important factors which affect
microbial xylanase synthesis are the usage of the transparent substrate
and potent microbes. Utilizing bountiful and fruitful farming residues
can enhance production and reduce the manufacturing cost in further
applications. The various research group studied Xylanases production
from different species by solid substrate fermentation and are listed in
Table 4.

Table 2
Pectinase production using various species and substrate.
Species Substrate Type of Fermentation pH Temperature References

Aspergillus niger LFP-1 Pomelo peel solid substrate fermentation pH 4.5 (30 ◦ C) Darah et al. (2015)
Bacillus subtilis Hazelnut shell submerged fermentation pH 7.0, 30 ◦ C Uzuner and Cekmecelioglu (2015)
Hydrolysate
Bacillus tequilensis wheat bran solid substrate fermentation pH 6 37 ◦ C, Chiliveri et al. (2016)
SV11-UV37
Aspergillus oryzae citrus waste and sugarcane bagasse Solid substrate fermentation pH 4 47 ◦ C Biz et al. (2016)
Rhizopus sp. C4 Peel of orange Solid substrate fermentation pH4-5 30 ◦ C Handa et al. (2016)
Bacillus subtilis SAV-21 orange peel and coconut fiber (4:1) Solid substrate fermentation pH 4.0 35 ◦ C Kaur and Gupta (2017)
Aspergillus fumigatus R6 rice bran Solid substrate fermentation pH4-5 33 ◦ C Wong et al. (2017)
Aspergillus niger orange pomace solid-state fermentation. pH 4.0 45–55 ◦ C Mahmoodi et al. (2017)
Bacillus sp. Y1 wheat bran Solid substrate fermentation pH 8.2 34 ◦ C Guo et al. (2019)
Aspergillus tamarii. Chikoo peels and pulp submerged fermentation pH (3–6) (30–60 ◦ C), Munir et al. (2019)
Aspergillus niger Biserrata leaves Solid substrate fermentation pH 4.0 50 oC Pagarra et al. (2019)
Aspergillus niger Nephroleis biserrata Solid substrate fermentation pH 4.0 50 oC Pagarra and Rahman (2019)
Aspergillus sp.VTMS Coffee Pulp Solid substrate fermentation pH (3–5) 30 oC Hidayah et al. (2020)
B. licheniformis KIBGE-IB3 Date fruit waste Solid substrate fermentation pH 7.0 37 oC Aslam et al. (2020)
Aspergillus niger Pineapple peel pectin Solid substrate fermentation pH 5.0 40 oC Ajayi et al. (2021)

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Table 3
Degumming of natural fibers by Pectinases produced from different species.
Species Enzyme Application Operating Observation Problems References
Conditions

Pseudozyma sp. SPJ Pectinase Flax fiber 12 h incubation Weight loss of 11 ± 1.2% No studies on Sharma et al.
(Yeast) degumming galacturonide release (2011)
Bacillus megaterium Pectate lyase Ramie fiber 24–48 h processing Nanoparticle supplemented Prolonged incubation Mukhopadhyay
AK2 (Bacteria) degumming time pectate lyase improves weight et al. (2013)
loss%
Bacillus tequilensis Pectate lyase, Poly- degumming of 300 U at 50 ◦ C in Weight loss, galacturonide acid Usage of chemicals Chiliveri et al.
SV11-UV37 galacturonase kenaf, sun hemp 11 h was higher with combined (2016)
(Bacteria) fiber +NaoH treatment methods
Aspergillus niger Pectinase Enzyme Banana Fiber 30 ◦ C incubation for Higher pectinolytic activity No studies on weight Sarma and Deka
Aspergillus extraction three days under solid-state, the tensile loss (2016)
fumigatus strength of fiber improved
(Fungus)
Fusarium oxysporum Pectin lyase Hemp fibers Glycine NaOH No evident study on fiber Usage of chemicals, Yadav et al. (2017)
MTCC 1755 treatment properties incubation time
(Fungi) + duration
24 h incubation
Bacillus cereus Pectinase Degumming of 10 h bacterial Pectin hydrolysis selective Weight loss was only Cheng et al. (2018)
ramie and kenaf treatment medium 4%
Bacillus Pectin lyase Flax degumming Degumming was Mixed system – advantageous, Weight loss was 6.33 Ge et al. (2019)
licheniformis, studied from 0h to higher degumming rate and 8.56%
Bacillus 160h
megatarium
Microbial Pectinase degumming 56h degumming + Ecofriendly, reuse of waste Prolonged incubation (Mao et al., 2019)
consortium process of ramie 0.2% NaOH residues increases economic time
RAMCD407 fiber treatment benefits
Bacillus cereus Calcium alginate degumming of 15 h, 45oC Maximum galacturonide No tensile strength Kohli and Gupta
immobilized and crude plant fibers released analysis (2019)
Pectin lyase

Some of the research findings are listed in Table 5.

4. Laccase

Laccases belong to the oxidases family and are widely found in a

diverse group of microorganisms. Production of Laccase from Mono­
cillium indicum was demonstrated with staging activity by Johannes and
Majcherczyk. (2000). The phenol-oxidase and ligninolytic property of
laccase was synthesized by Pycnoporus cinnabarinus and Pycnoporus
sanguineus, respectively. The three-dimensional image of the Laccase
enzyme is illustrated below in Fig. 3.

4.1. Production of Laccase by fermentation

Both solid-state and submerged fermentation methods produced the

Fig. 2. The 3-D image of Xylanase enzyme.
3.2. Application of xylanases in degumming
Fibers can be degummed by using xylanolytic enzymes with pecti­
nolytic enzymes (Puchart et al., 1999). Even though pectinases pursue a
potent role in degumming, the involvement of xylanase is also consid­
ered (Cao et al., 1992). The conventional slow retting process can be
replaced by enzymatic methods, creating a platform for new fiber
liberation technology. The enzymatic degumming creates less damage to
the cellulosic substance of the natural fiber. Xylanases play a major role
in the degumming of ramie fibers. Extensive research has been carried
out on the degumming of various natural fibers by Xylanaces produced
from different species. The fibers with xylanase treatment were found to
possess high linear density fibers with low prices compared to the Lac­
case treatment as it ends up with lower linear density and high cost.
Laccase enzyme. Solid substrate fermentation has advantages over
submerged fermentation in yield rate, waste production, lower energy
needs, cost-effective media. Literature studies suggest solid-state
fermentation has immense application in synthesizing antibiotics, bio­
cides, surfactants, and ligninolytic enzymes by using agricultural residue
as substrates. It can be adopted by inert solid support for easier recovery
of extracellular products and is cost-effective (Rodríguez Couto and
Sanromán, 2019). Many researchers have synthesized Laccase enzymes
using different species and substrates and are listed in Table 6.
4.2. Application of laccases in degumming
Laccase enzyme will deteriorate the dense, sticky material in the
interior core and lateral surface, producing cellulose-rich fibers. Lac­
cases are reaction-specific catalysts with focused performance, inex­
pensive, and environment-friendly, leading to the acceptance of laccase
in degumming applications. Laccases are potently capable of lignin
degradation, and therefore it can be accomplished in bast fibers
degumming. The potential of laccase enzymes in refining bamboo fibers,
enhanced lignin removal was successfully reported by Liu et al. (2012).
They also highlighted bamboo fibers refining xylanase and laccase with
limited loss of strength (Sreenivas et al., 2020). George et al. (2016)

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Table 4
Xylanases production by various species using different substrates.
Species Substrate Type of Fermentation pH Temperature References

Aspergillus niger DFR-5 Mixed fruit peel waste Solid substrate 5.0 37 C
◦
Pal and Khanum (2010)
fermentation
Bacillus arseniciselenatisDSM 15340 Wheat bran Solid substrate 8.0. 50 ◦C Kamble and Jadhav
fermentation (2012)
Aspergillus fumigatus SK1 Palm oil waste Solid substrate 4.0 70 ◦ C Ang et al. (2013)
fermentation
Mucorindicus, Mucorhiemalis, and Rhizopus oryzae Wheat bran Solid substrate 4–5 40.0, 43.4, and 43.4 ◦ C, Behnam et al. (2016)
fermentation respectively.
Aspergillus niger, Aspergillus oryzae, and Aspergillus Wheat bran Solid substrate 4.0 Yin et al. (2018)
awamori fermentation
o
Endophytic fungi Lignocellulosic Solid substrate 4.5–6 30–70 C Marques et al. (2018)
substrates fermentation
Aspergillus nigerCCUG33991 waste of corn, bran Solid substrate 4–5 28–35 oC Khanahmadi et al.
fermentation (2018)
Trichoderma asperullum UC1 Raw oil palm frond Solid substrate 3.0 30 ◦ C Ezeilo et al. (2019)
leaves fermentation
Rhizopus oryzae UC2 Raw oil frond leaves Solid substrate 3.0–5.0 50–60 oC Ezeilo et al. (2020)
fermentation
Aspergillus niger Rice husk Solid substrate 5.0 30 ◦ C Afiqah Razali et al.
fermentation (2020)
Aspergillus niger Corn hub and saw dust Solid substrate 3.0 45 ◦ C Ire et al. (2021)
fermentation

Table 5
Degumming of natural fibers by xylanases using different species.
Species Enzyme Application Conditions Observation Problems /Disadvantage References

Bacterial consortium Xylanases Screening of ramie- Diversity, characterization Pectinase was the key Some strains produced Duan et al.
pectinases degumming strains of degumming strains enzyme of ramie cellulase and degraded (2012)
degumming, cellulose
Bacillus sp. HG-28 Xylanases Ramie fiber 16h microbial degumming Efficient than chemical Usage of chemicals, cellulase Fan et al.
Pectinase + degumming, gum content production (2015)
NaOH treatment reduction
Bacillus cereus P05 Xylanases Ramie fiber 60 h retting process Breaking strength, gum Prolonged retting time Wang et al.
Pseudomonas sp. pectinases removal increased (2017)
X12
bacterial consortium Xylanases ramie degumming 56 h followed by 0.2% Recovery of pectin crystals Usage of chemicals, cellulase Mao et al.
RAMCD407 pectinases NaOH treatment. and xylan chips production (2019)

Fig. 3. The 3-D image of Laccase enzyme.

reported the fast growth of enzyme application in natural fiber modifi­ analytical results of degummed natural fibers. The analytical results
cation. Some of the observations on degumming of fibers by Laccases are included in our study are tensile analysis, lignin quantification, hemi­
listed in Table 7. cellulose and cellulose content, density, moisture, and ash content.

5. Analysis of degummed natural fibers

Degummed natural fiber exhibits modification in the properties and
composition of fibers. Efficient degumming results in the significant
removal of hemicellulose substances. The hemicellulose removal can be
confirmed by analyzing the chemical composition of natural fibers
before and after degumming. In our study, we have illustrated the
5.1. Determination of tensile strength
Tensile strength measures the breaking strength, force resistance,
and yield strength of the natural fibers. It can be analyzed using tensile
identifying equipment under the ASTMD3822 standard. Microwave
pretreatment was experimented with hemp fibers to remove the hemi­
cellulose substance. The tensile strength of degummed hemp fiber was

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Table 6
Laccase enzyme production by various species and substrates.
Species Enzyme Substrate Fermentation Reference

Trametes vesicolor Crude laccase Agro residues Solid substrate Stoilova et al. (2010)
fermentation
Lenintu laedodas Laccase and endoglucanase Reed grass, beanstalks, wheat Solid substrate Philippoussis et al. (2011)
production straw fermentation
Coriolopsis caperata RCK2011 Laccase Corn steep liquor Solid substrate Nandal et al. (2013)
fermentation
Aspergillus species Laccase Winery and olive mill waste Solid substrate Salgado et al. (2014)
fermentation
Pleurotus ostreatus laccase Sugarcane bagasse Solid substrate Karp et al. (2015)
fermentation
Pseudolagarobasidium acaciicola Laccase production Parthenium biomass Solid substrate Adak et al. (2016)
LA1 fermentation
Pleurotus ostreatus laccase and peroxidase of and lignin Potato peel waste Solid substrate Ozcirak and Ozturk (2017)
fermentation
Pleurotus eryngii Laccase Peach waste Solid substrate Akpinar and Ozturk (2017)
fermentation
Trametes versicolor IBL-04 Alginate-chitosan Immobilized corncobs Solid substrate Asgher et al. (2018)
Laccase fermentation
Ganoderma lucidium Laccase Wood waste Both fermentation Rodrigues et al. (2019)
Trametes versicolor Laccase Tea residue Solid substrate Xu et al. (2020)
fermentation
Galactomyces geotrichum Laccase Olive leaves Solid substrate Pourkhanali et al. (2020)
fermentation
Halopiger aswanenesis Lignolytic enzymes Wheat bran Solid substrate Chauhan and Choudhury
fermentation (2021)
Pycnoporus sanguineus Laccase – Solid substrate Hafid et al. (2021)
fermentation

Table 7
Degumming of fibers by Laccases.
Process Enzyme Application Observation References

Laccase mediator system
treatment
Enzymatic treatment
Laccase and TEMPO
mediated system
Chemo-enzymatic
modification
Enzymatic treatment
Laccase Changes of lignin in the Improves mechanical properties, surface Zhang et al. (2014)
jute fiber hydrophobicity
Laccase, Xylanase Study on banana fiber Lignin, hemicellulose removal was higher Vishnu Vardhini and
properties Murugan (2017)
Laccase and Grafting of silk fiber The concentration of amino group decreased, Zhou et al. (2017)
2,2’tetramethylpiperidine-N-oxyl self cross-linking of fibers
Laccase Hemp fiber degumming Improves moisture absorption, water retention Yeping et al. (2019)
value,
Laccase, Pectinase Banana fiber Removal of pectinolytic substances, surface Paramasivam et al. (2020)
degumming smoothening of banana fiber

increased to 12.66 cN from 10.26 cN (Han et al., 2011). The Peroxide
treatment of ramie fibers possesses a moderate decrease in the length,
tenacity, and elongation property. The decreased tenacity corresponds
to the breakage of microstructures and cellulosic fibrils (Li and Yu,
2014). Degumming banana fiber with the steam explosion method
resulted in higher breaking strength of 156.37 Mpa (Sheng et al., 2014).
An increase in the concentration of alkali in degumming of lotus fibers
exhibits a decrease in the reduction of braking force from 5.31 cN to
2.37 cN and reduced elongation percentage (Li and Fu, 2015). The
oxidized ramie fibers have increased tenacity to 6.18 cN/dtex (centi­
newton decitex) from 3.02 cN/dtex (centinewton decitex). The increase
in tenacity is due to the successful removal of non-cellulosic substances,
delignification of hemicellulose, and pectin degradation.
Further, the tensile property of fiber is directly proportional to the
degree of polymerization. The degummed ramie fiber has an inclined
polymerization degree. The increase in tenacity and polymerization
degree of ramie fiber was due to the degradation of non-cellulosic chains
to smaller compounds (Li et al., 2015). Meng et al. (2017) reported that
the alkali coupled with magnesium hydroxide treated ramie fiber has
resulted in the tenacity and elongation of ramie fiber. The substitution
rate of magnesium hydroxide above 20% in the degumming process
causes a decrease in the tensile strength of the ramie fiber. Enzymatic
treatment assists the removal of fragile structures and modifies the
molecular behavior of fibers. The modification in molecular behavior
corresponds to the amplified tensile strength compared to the chemical
degumming methods (Balakrishnan et al., 2019).
5.2. Determination of lignin, hemicellulose, and cellulose content
The major parameters that influence the chemical composition of
fibers are the locality of the plant, maturity, degumming methods, and
characteristics of the fiber. The matrix formation of cellulose and
hemicellulose was due to the presence of non-cellulosic layers. The
lignin content corresponds to the rigidity of the plant. Li and Yu. (2014)
reported that the per-oxide treatment of ramie fibers has successfully
reduced the hemicellulose content from 14% to 5%. But the peroxide
treatment was less efficient in lignin removal of ramie fibers. The
oxidized ramie fibers have increased cellulose content from 73% to 95%
and decreased non-cellulosic content from 13% to 3%. The degree of
gummy substance removal is reflected in the reduction of residual gum
content. The efficient pectin and non-cellulosic substance removal lead
to finer and separated cellulosic fibers (Li et al., 2015). The peracetic
acid-treated jute fiber has increased cellulose content, reduced hemi­
cellulose, and lignin content. The reaction of the lignin with peracetic
acid causes electrophilic hydroxylation of aromatic rings, oxygenated
groups. The hydroxylation results in the formation of quinone, lactone,
and muconic acid and demethoxylation (Duan et al., 2017). Jiang et al.
(2018) demonstrated that the green degumming of ramie fibers results

6
M. Subash and P. Muthiah
in the efficient removal of the gummy substance from 27% to 5%.
Further, green degumming has improved the fineness of ramie fiber to
1400Nm and also increased the whiteness of fiber to 50%. The
degumming of kenaf fiber has significantly removed hemicellulose and
lignin content. Zhang et al. (2019) observed that the novel degumming
of kenaf fiber has also increased the percentage of cellulose content and
absorption rate.
5.3. Determination of density and diameter
ASTM D 1505-03 is the standard method to measure the density of
the fibers. Linear density can be calculated using ASTM D 1577-07.
Solvents used for the determination of density are xylene and carbon
tetrachloride. Fiber fineness can be measured in Tex (weight in grams
per unit length of fiber). The efficient removal of hemicellulose sub­
stance corresponds to an increase in linear density of fiber and finer
fibril molecules (Devireddy et al., 2018).
5.4. Determination of moisture and ash content
The moisture of natural fiber will be measured and expressed as a
percentage. The oven conditions the fiber samples at 105oC, and the
weight (WI) will be determined before and after conditioning as per
ASTM D 2495-07 standard. The ash content determines the inorganic
residues after heating at 600oC for 4–5 h. The banana fibers were sub­
jected to the steam explosion at 800oC. The ash content of banana fiber
was reduced to 1.72 ± 0.12 from 7.02 ± 0.32 by degumming the banana
fiber with a steam explosion (Sheng et al., 2014). Li and Yu (2014) re­
ported that the ramie fibers were degummed by alkali, peroxide, and
isopropyl alcohol coupled with peroxide and compared with control
ramie fibers. The ash content of control, alkali, peroxide, peroxide
associated with isopropyl alcohol were 2.45%, 1.42%, 1.02%, and
1.24%, respectively(Li and Yu, 2014). The oxidation-reduction potential
on the degumming of ramie fibers with hydrogen peroxide has
decreased the ash content of ramie fibers from 2.45% to 0.28% (Li and
Yu, 2015). Similarly, Song et al. (2019) reported that the combined
steam explosion and chemical degumming of kudzu fiber had reduced
the ash content.
5.5. Weight loss analysis
Weight loss analysis is done to identify the gum loss of control and
degummed natural fibers. The weight loss of degummed fibers can be
calculated concerning the initial weight of the fibers. Many researchers
highlighted the advantages of enzymatic degumming in significant
weight loss. Ibrahim et al. (2019) reported that the scouring of cotton
fabrics with polygalacturonase enzyme resulted in 5.2% weight loss
(Ibrahim et al., 2019). The gum content was reduced to 11.6% from
25.9% on enzymatic degumming of ramie fiber (Jiang et al., 2018). The
literature studies show that enzymatic treatment of jute fiber contributes
to a weight loss of 12–17%. Similarly, the cotton fabric treated with
enzymes also resulted in a weight loss of 10% (Rajulapati et al., 2020).
The weight loss percentage of enzymatic degummed fiber was signifi­
cant compared to conventional degumming methods.
6. Characterization of degummed fibers
Characterization studies can examine the surface modification of
fibers. The characterization studies reported in this research include
Microscopy analysis, SEM, FESEM, ESEM, FTIR, XRD, NMR, and thermal
analysis. Characterization studies will examine degummed fibers to
identify the pros and cons of degumming methods.
6.1. Microscopy analysis
An optical microscope, commonly known as a Light microscope, is
7
Cleaner Engineering and Technology 5 (2021) 100304
used to visualize the magnified view of the sample through the series of
lens magnification under visible light. Polarized light microscopy is one
of the members of the optical microscope by involving the illumination
of the sample under polarized light. A fluorescence microscope is also a
member of the optical microscope, which employs not only fluorescence
but also few features such as scattering, reflection, attenuation, and
absorption. Microbial retting of ramie fibers exhibits an average diam­
eter of 32um, greater than chemically treated rame fiber (Angelini et al.,
2015). The mechanically extracted banana fiber was comparatively
rough and less significant under the light microscope (Balakrishnan
et al., 2019). For a light microscopic view of ramie fibers, bast tissue,
gum debris, and treated ramie fiber were placed on a microscopic slide
with the aid of tweezers and a drop of distilled water. For a Polarized
microscopic view of ramie fibers, samples were illuminated with a
mercury lamp for 212 milliseconds. The reference of ramie xylan, pectin,
and lignin under illumination was an indigo blue image, dark orange,
and red image, respectively. The replacement of reference color with
bright white infers the removal of non-cellulosic substances (Mao et al.,
2019). The Autofluorescence signal of the non-cellulosic ramie gum
component is visualized by the dark orange fluorescent signal emitted
by ramie pectin particles. Similarly, the ramie xylan particles emit bright
indigo fluorescent signals, and lignin emit a red fluorescent signal.
6.2. Morphological surface studies
Scanning Electron Microscopy generates signal diversification at the
outer layer of the solid specimen by high-energy electrons with a focused
beam. Generated signals produce a 2-dimensional image that reveals
knowledge of sample texture and chemical composition. Comparative
analysis of the surface of sample fibers with control can be examined
using a scanning electron microscope. Removal of gum material from
the exterior of the bast fiber cell and their modification into single bast
fibers can be examined by SEM image (Beltran et al., 2002). Li and Fu
(2015) demonstrated that the alkali degumming of lotus fibers results in
the removal of binding agents without damaging the nature of the fibers.
Oxidation degumming of ramie fiber has resulted in fine pits and pla­
ques, thereby losing fibrous structures (Li et al., 2015). Duan et al., 2017
reported that the SEM image of acid-treated Jute fiber had shown the
partial removal of gummy substance with few elementary fibrils in the
primary layer of jute fiber. Researchers also reported that most gummy
substances were detached from the exterior of the ramie fiber bundle by
microbial degumming of ramie fibers. The SEM images of degummed
ramie fiber surface were clear and smooth (Mao et al., 2019). Similarly,
research on degummed kenaf fiber also resulted in reduced gum mate­
rials compared to raw kenaf fibers. Further, literature studies also infer
that green degumming, such as microbial and enzymatic degumming of
ramie fiber, had comparable efficiency to the conventional fiber
degumming analysis (Jiang et al., 2018).
6.3. Field emission scanning electron microscope
The field emission scanning electron microscopy (FESEM) is
employed to visualize the topographic details of a sample surface,
fractionated objects, nanomaterials, and nanocomposites. FESEM is one
of the types of electron microscope which scans the sample surface with
a high-energy beam of electrons in a raster scan pattern. Rajulapati et al.
(2020) reported that the FESEM image of microbially degummed jute
fibers, chemical degummed jute fibers, and control jute fibers were
compared. The microbially degummed jute fibers exhibit light brown
color, smoother surface, removal of pectin and wax. Whereas, chemi­
cally treated jute fibers and control fibers were in yellow and dark brown
color respectively. Similarly scouring of cotton fabrics was also analyzed
by FESEM. The image of enzymatically treated cotton fabrics, chemi­
cally treated cotton fabrics, and control fabrics were light
yellowish-white, whitish, and yellowish-white. The smoothness on the
surface of enzyme-treated cotton fabrics was efficient compared to other
M. Subash and P. Muthiah
methods(Rajulapati et al., 2020).
6.4. Environmental scanning electron microscopy
Environmental scanning electron microscopy (ESEM) is a unique
technique that involves the examination of uncoated biological material
and industrial materials with a high electron beam in a high chamber
pressure atmosphere of water vapor. The major applications of ESEM are
dynamic experiments, drying, and crystallization. ESEM employs a
scanned electron beam, electromagnetic lenses for the electron beam’s
direction on the specimen’s surface. The interaction of specimen surface
and electron beam results in various signals collected with appropriate
detectors. The ESEM micrographs of electrospun silk fibers were circular
in cross-section and smooth surface. The electrospun fibers were spun
with a 3 kV/cm (Ayutsede et al., 2005). Li and Fu (2015) reported that
the lotus fibers plucked at different growth stages were examined by
ESEM analysis. The surface of the control lotus fibers was coarse, con­
nected by pectin to form fiber bundles. The surface of degummed lotus
fibers was individual in longitudinal view with the removal of gummy
substances. The ESEM image of enzymatically treated flax fibers showed
significant sticky substance removal, leading to the formation of a single
fiber (Xiang et al., 2020).
6.5. Functional group analysis
The physiochemical properties of the lingo cellulosic materials can
be investigated by a non-destructive method. FTIR analysis can be
executed to examine the modifications in the spectral regions of bast
fiber structures before and after treatment (Beltran et al., 2002). The
intensification of cellulose composition can be observed by the sharp­
ening peaks attributed to the fibers’ crystallinity (Cherian et al., 2008).
The height at 1,735cm-1 corresponds to the carboxylic esters in pectin
and wax. Lignin usually peaks at 1,510 cm-1 which disappears on
degumming of Hemp fibers (Han et al., 2011). Microwave-assisted
degumming of hemp fibers exhibits the disappearance of the lignin re­
gion, unchanged cellulose region, and presence of glycosidic bonds (Han
et al., 2011). The spectrum of peroxide treated ramie fiber exhibits
higher C–H stretching, CH2 symmetric bending, and C–O stretching
intensities. The higher intensities correspond to the successful removal
of gummy substances (Li and Yu, 2014). The oxidation degumming of
ramie fibers causes breakage in the hydrogen bond structures.
Further, increased oxidation rate causes increased stretching vibra­
tions, thereby leading to aldehyde and carboxyl groups (Li et al., 2015).
The alkali degummed ramie fiber research highlighted the efficiency of
magnesium hydroxide in degumming relies on the substitution rate. The
substitution of magnesium hydroxide above 20% affects the cellulosic
nature of the fibers (Meng et al., 2017). Degummed jute fiber was found
to have reduced peak at 2,900 cm-1 and 1,420 cm-1 indicating the
removal of hemicellulose and lignin (Duan et al., 2017). FT-IRM analysis
of degummed kenaf fiber shows the breakage of hemicellulose sub­
stance, making the surface of the kenaf fiber smooth and clean. The
researcher also highlighted that microbial degumming was more effi­
cient in removing a non-cellulosic substance from the kenaf fiber (Zhang
et al., 2019). Green degumming of ramie fiber effectively removed the
lignin in degummed ramie fibers (Jiang et al., 2018). The alkali
degumming of lotus fibers disturbs the hydrogen bond of cellulose and
also cause substitution of sodium atom in place of hydrogen. The
ATR-FTIR analysis of enzymatic degummed jute fibers possesses sig­
nificant change in alkane and alcohol groups of the pectin and also loss
of methylation groups in the pectin region (Rajulapati et al., 2020)
6.6. Thermogravimetric analysis
The thermal analysis of the fibers is done to identify the reduction in
weight of the fiber as functional temperature. T-curve provides infor­
mation about the thermal stability, composition of the initial sample,
8
Cleaner Engineering and Technology 5 (2021) 100304
intermediate products, and composition of the solid residue. Sheng et al.
(2014) outlined the higher ash content of the raw banana fiber
compared to the treated banana fiber at 800oC which highlights the
elimination of lignin. To analyze the thermal properties of silk fibers,
sample fibers were placed in aluminum pans and weighed using a mi­
crobalance. The thermal property of the sample was analyzed by a
thermal analysis system using nitrogen as flushing gas. The temperature
was varied from 25oC to 800oC at a rate of 10 oC/min. Derivative
thermogravimetric analysis (DTA) is carried out to find the rate of
weight loss of the material on heating. In thermogravimetric analysis,
the derivative of the TG curve concerning time is directly correlated with
the differential equations of the rate of weight change of the degummed
fiber. The variation in thermostability of the sample was studied by
thermogram and differential thermogram (Wang et al., 2015). The
thermal analysis of degummed ramie fiber exhibited weight loss be­
tween 300 and 364oC. The weight loss was due to the depolymerization
of hemicellulose, pectin, and glycosidic bond cleavage. The lignin
decomposition of degummed ramie plant occurs between 367–776oC
(Kandimalla et al., 2016). Depolymerization of non-cellulosic substances
and the glycosidic separation of cellulose and carbon residues are car­
ried out while subjecting to a temperature between the range of 200 and
300oC (Devireddy et al., 2018). An increase in degumming temperature
corresponds to the increased degumming capability and efficient gum
removal (Guoliang et al., 2019). Degumming of bast fibers results in the
decomposition of amorphous cellulose. The subtle decrease in thermal
stability does not affect the quality of bast fibers (Song et al., 2019).
6.7. X-ray diffraction analysis
Natural fibers contain lignin, hemicellulose, and alpha-cellulose. The
nature of cellulose and lignin are crystalline and amorphous, respec­
tively. An increase in crystallinity indicates the increase in the compo­
sition of cellulose in the fibers. Han et al. (2011) observed that the
crystallinity of degummed hemp fiber was increased to 85% from 80%.
The per-oxide treatment of ramie fibers has increased crystallinity.
Increased crystallinity corresponds to the decrease in the softness of the
fibers. The per-oxide treatment of ramie fiber produces higher cellulosic
content by the efficient removal of amorphous compounds (Li and Yu,
2014). Sheng et al. (2014) reported that the percentage of crystallinity is
53.9% for enzymatically treated banana fiber which is comparatively
higher concerning the control sample with 11.8%. This statistic data
proves the treated banana fiber shows a higher crystalline nature. The
oxidation degumming of ramie fibers possesses higher crystallinity of
about 78.91% due to the efficient neglection of amorphous
non-cellulosic compounds. The absorption peaks of degummed ramie
fiber have a higher intensity than the control ramie fibers (Li et al.,
2015). The alkali degumming of lotus fibers taken from all the growth
stages possesses crystallinity (Li and Fu, 2015). The crystallinity of the
degummed jute fiber was increased from 65.27% to 82.02%. The rapid
increase in crystallinity of degummed jute fiber was due to the efficient
removal of hemicellulose substances (Duan et al., 2017). The alkali
degummed ramie fiber was found to have a higher crystalline plane
family. The alkali coupled with NaOH and Mg(OH)2 causes lower
crystallinity and the presence of few amorphous residual gums (Meng
et al., 2017).
6.8. Van Soest (detergent fiber analysis)
The principle of Van Soes is the separation of plant cell wall com­
ponents into less digestible components (such as hemicellulose, lignin,
and cellulose) and mostly digestible components such as starch and
sugars. The detergents used to separate cell wall components are Na-
lauryl sulfate, Ethylene diamine tetraacetic acid, and Cetyl trimethyl
ammonium bromide. Comparative analysis on ramie fibers’ chemical
and microbial degumming infers that most of the hemicellulose sub­
stances have been removed by chemical degumming (Angelini et al.,
M. Subash and P. Muthiah
2015).
6.9. Nuclear magnetic resonance (NMR)
Nuclear Magnetic Resonance Spectroscopy is an analytical technique
to measure the quantity of each compound in a sample mixture. It is a
technique used in quality control to determine the molecular structure of
the mix. Researchers have adopted NMR Spectroscopy analysis of
degummed jute fiber to examine the modification like cellulose due to
the acid degumming process. The degummed jute fiber possesses C-6
crystalline cellulose, amorphous cellulose, C-4 interior cellulose, and
crystal surface cellulose. Apart from the crystalline nature, the control
and treated jute fiber structure found variation in the supermolecular
form. Duan et al., 2017 observed that the amorphous cellulose of the
acid degummed jute fiber was partially defected due to the acid
treatment.
7. Conclusion
Textile industries face major challenges in improving the wet textile
processing of synthetic fibers. The negative impacts of synthetic fiber
processing are water consumption, energy consumption, and harmful
chemical effluents. The present study exploits environmentally benign
and sustainable alternative fibers such as jute, ramie, hemp, bast, and
banana fibers. Among the natural fibers, very limited research analysis
has been reported on the enzymatic processing of banana fibers. Enzy­
matic degumming stands as a positive approach and also replaces
drawbacks of chemical degumming. Enzymatic degumming of natural
fibers exhibits efficient removal of a non-cellulosic substance without
altering the cellulosic content. Literature studies suggest that efficient
degumming of fibers indicates significant gum weight loss, improved
tensile strength. The proliferation in cellulose content corresponds to the
crystallinity of the fibers, which can be analyzed by the characterization
of the degummed fibers with surface morphology, functional groups,
thermal analysis, and other quantitative estimation of fiber contents.
Enzymatically degummed natural fibers provide a remarkable change in
the textile industries as it is eco-friendly and sustainable.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
CRediT authorship contribution statement
Mira chares Subash: Conceptualization, Methodology, Data cura­
tion, Writing – original draft, Writing – review & editing. Perumalsamy
Muthiah: Conceptualization, Methodology, Supervision, Writing – re­
view & editing.
Acknowledgment
The authors gratefully acknowledge the Department of Biotech­
nology (DBT) for the financial assistance through Biotech Consortium
India Limited (BCIL) with sanctioned reference number (DBT-NER/
AGRI/33/2016 ft. March 22, 2016 of DBT & BCIL/NER-BPMC/2018/
245 ft. March 26, 2018 of BCIL) is kindly acknowledged.
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