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Characterisation of α-chitin extracted

from a lichenised fungus species
Xanthoria parietina
ab c c
Murat Kaya , Mehmet Gökhan Halıcı , Fatih Duman , Sevil
d e
Erdoğan & Talat Baran
a
Department of Biotechnology and Molecular Biology, Faculty of
Science and Letters, Aksaray University, 68100 Aksaray, Turkey
b
Science and Technology Application and Research Center,
Aksaray University, 68100 Aksaray, Turkey
c
Click for updates Department of Biology, Faculty of Science, Erciyes University,
38039 Kayseri, Turkey
d
Fisheries Programme, Ke¸an Vocational College, Trakya
University, 22800 Ke¸an, Edirne, Turkey
e
Department of Chemistry, Faculty of Science and Letters,
Aksaray University, 68100 Aksaray, Turkey
Published online: 02 Jan 2015.

To cite this article: Murat Kaya, Mehmet Gökhan Halıcı, Fatih Duman, Sevil Erdoğan & Talat Baran
(2015): Characterisation of α-chitin extracted from a lichenised fungus species Xanthoria parietina,
Natural Product Research: Formerly Natural Product Letters, DOI: 10.1080/14786419.2014.995651

To link to this article: http://dx.doi.org/10.1080/14786419.2014.995651

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 Natural Product Research, 2015
http://dx.doi.org/10.1080/14786419.2014.995651

SHORT COMMUNICATION
Characterisation of a-chitin extracted from a lichenised fungus species
Xanthoria parietina
Murat Kayaa,b*, Mehmet Gökhan Halıcıc, Fatih Dumanc, Sevil Erdoğand and Talat Barane
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a
Department of Biotechnology and Molecular Biology, Faculty of Science and Letters, Aksaray University,
68100 Aksaray, Turkey; bScience and Technology Application and Research Center, Aksaray University,
68100 Aksaray, Turkey; cDepartment of Biology, Faculty of Science, Erciyes University, 38039 Kayseri,
Turkey; dFisheries Programme, Keşan Vocational College, Trakya University, 22800 Keşan, Edirne,
Turkey; eDepartment of Chemistry, Faculty of Science and Letters, Aksaray University, 68100 Aksaray,
Turkey
(Received 2 July 2014; final version received 30 November 2014)

Lichens are symbiotic associations formed mainly by ascomycete fungi and green
algae or cyanobacteria. The presence of chitin in the fungal cell wall has been revealed
by previous studies. Considering the presence of fungi in the lichens, this work
determines the presence of chitin in a cosmopolitan lichen species Xanthoria parietina.
In this study, chitin was derived from a lichen species for the first time and its
physicochemical properties were determined by Fourier transform infrared
spectroscopy, thermogravimetric analysis, X-ray diffraction, scanning electron
microscopy and elemental analysis. The dry weight chitin content of X. parietina
was 4.23%, and this chitin was in the a-form. The crystalline index value of the lichen
chitin was calculated as 70.1%. The chitin from X. parietina had a smooth surface.
Keywords: lichen; chitin; isolation; characterisation; biopolymer

1. Introduction
Chitin is a biopolymer that can be used for various purposes, such as in textiles as well as in the
biomedical and food industries (Dutta et al. 2004; Rinaudo 2006). Although there are millions of
organisms that contain chitin in their body, only certain crustaceans (crayfish, prawn, shrimp,
crab, krill and lobster) are used to produce commercial chitin (Dutta et al. 2004; Rinaudo 2006).
Apart from these organisms, insects, fungi, sponges and corals have been investigated as chitin
sources recently (Ifuku et al. 2011; Bo et al. 2012; Liu et al. 2012; Ehrlich et al. 2013). In this

*Corresponding author. Email: muratkaya3806@yahoo.com

q 2015 Taylor & Francis

 2 M. Kaya et al.

study, the authors preferred widely distributed species belonging to lichens that have not been
investigated previously in terms of chitin content.
Previous studies on lichens have focused on the systematics, ecology, distribution and
molecular biology (Grube et al. 1995; Hawksworth et al. 2011). In recent years, many studies on
secondary chemicals such as depsides, depsidones, dibenzofurans, anthraquinones and
xanthones produced by lichens and fatty acid composition, cytotoxic, antimicrobial, anticancer
and antioxidant properties of lichens have been carried out (Tanahashia et al. 2001; Matheya
et al. 2002; Vila et al. 2011; Masters & Bräse 2012; Kosanic et al. 2013). Unfortunately, there
have been a limited number of studies related to the chitin of the 20,000 lichen species
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worldwide (Dahlman et al. 2002; Meichik & Vorob’ev 2012).

Lichens are symbiotic organisms comprising a fungus and a green algae or cyanobacteria.
Lichens can be found in some of the most extreme environments on earth, and most lichens are
sensitive to air pollution (Dobson 2000). The material used in this study, Xanthoria parietina
(Ascomycota, Teloschistaceae), is a common epiphytic lichen that especially lives in nitrogen-
rich habitats, such as villages where agricultural activities are abundant. It has a foliose thallus
that is bright orange, particularly in sunny habitats, and it is one of the lichens that can be
collected relatively widely, when compared with other lichens.
This study aimed to isolate chitin, for the first time, from a lichen species, and determined the
dry weight chitin content as well as whether this lichen can be used as a chitin source for current
chitin needs. In addition, it also aimed to determine the physichochemical properties of the
lichen chitin and to compare them with the properties of chitins obtained from crab, prawn,
shrimp, insects and fungi in previous studies.

2. Results and discussion

2.1. Chitin content in X. parietina
In this study, chitin was obtained from a lichen species X. parietina, and it was determined that
the dry weight chitin content in this lichen species was 4.23%. The chitin content of X. parietina
is less than the chitin contents (10 – 40%) of the shell structures from organisms such as crab,
crayfish, prawn and shrimp (Tolaimate et al. 2003; Liu et al. 2012). Despite this, it is possible
that X. parietina can be an important chitin source, considering that it is a cosmopolitan species
that is found in large amounts, which can be easily collected. Previous studies showed that the
dry weight chitin contents of fungi vary between 1% and 20%, depending on the fungi species
and methods used (Vetter 2007; Di Mario et al. 2008; Ifuku et al. 2011). It was observed that
glucan could not be removed completely from the fungal chitin using chemical isolation
methods (Di Mario et al. 2008; Ifuku et al. 2011).

2.2. FT-IR
When previous studies were examined, it was observed that the amide I band in the FT-IR
spectra of the chitin in the a-form was split into two different bands at 1660 and 1620 cm21.
It was reported that the amide I band in the FT-IR spectra of b chitin gave a single peak around
1620 cm21 (Gardner & Blackwell 1975). Considering the FT-IR spectra of the chitin obtained
from X. parietina in this study, it was observed that amide I gave two bands at 1654 and
1624 cm21, and the presence of these bands showed that the obtained chitin was in the a-form.
In addition, the presence of bands at 3430, 3268, 3101, 2919, 2853, 1560, 1422, 1375, 1310,
1153, 1110, 1060, 1012, 954 and 891 cm21 in the spectra is another indication of a-chitin
(Acosta et al. 1993) (Figure S1).
In this study, the FT-IR peaks and spectra of the chitin obtained from X. parietina and
commercial shrimp chitin bought from Sigma-Aldrich (Pcode: 1001416772) are shown in
 Natural Product Research 3

Table S1 and Figure S1. As seen from the figure and table, the FT-IR analysis results of the
lichen chitin are quite similar to those of commercial chitin, which confirms that chitin can be
obtained from lichen species.

2.3. Thermogravimetric analysis

According to the results of the thermogravimetric (TGA) analysis of the chitin obtained from
X. parietina, mass loss occurred in two stages (Figure S2). In the first step, the mass loss of 5%
between 25 and 1508C was probably due to the evaporation of water in the chitin. Another mass
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loss (61%) was observed between 150 and 6008C in the second step, and it can be attributed to
the decomposition of the chitin. The maximum decomposition temperature (DTGmax) of the
lichen chitin was recorded as 3428C. Considering the TGA analysis results of a-chitins obtained
from organisms such as prawns, crabs, shrimp and insects in earlier studies, the mass losses were
also observed in two stages, similar to this study (Al Sagheer et al. 2009; Sajomsang & Gonil
2010). According to Jang et al. (2004), while the DTGmax value of b-chitin is around 3008C, for
a-chitins this value is around 3508C. In this study, the DTGmax value showed that the lichen
chitin was in the a-form. These results agree with earlier studies.
Considering the TGA analysis results of commercial chitin, mass loss was observed in two
steps as in the lichen chitin (Figure S2(b)). The DTGmax value (3868C) of commercial chitin
recorded was higher than that from lichen chitin. In addition, the percentage mass loss (75%) in
the second decomposition step of the commercial chitin was found to be higher than that in the
lichen chitin. As a result, it was revealed that the lichen chitin has lower thermal stability than
commercial chitin. This probably stems from glucan residues that cannot be fully removed from
the lichen chitin.

2.4. X-ray diffraction

A total of seven peaks (9.08, 12.3, 19.2, 20.8, 23.64, 26.64 and 27.48) were observed (Figure S3)
in the X-ray diffraction (XRD) analysis. Of these, 9.08, 19.2, 26.64 and 27.48 were sharp peaks,
while 12.3, 20.8 and 23.648 were weak peaks (Figure S3). The crystalline index (CrI) value of
the lichen chitin was calculated as 70.1%. It has been observed that the CrI values of chitins
extracted from the shells of animals such as crab and shrimp varied between 80% and 90%
(Sajomsang & Gonil 2010), while the CrI values of chitins obtained from some fungal species
varied between 47% and 80%, depending on the species (Ifuku et al. 2011). The CrI values of
fungal chitins are lower than those of chitins obtained from animals such as crab and
shrimp. This situation stems from the b-1,3 glucan, which cannot be fully removed from the
chitin during the isolation procedure (Ifuku et al. 2011). Considering the CrI value determined in
this study, it is possible that there were glucan residues in the obtained chitin sample.
There were small differences in terms of XRD analysis results between commercial chitin
obtained from shrimp (Sigma-Aldrich, Pcode: 1001416772) and lichen chitin in this study. The
peak at 26.648 in the lichen chitin was quite sharp compared to that in the commercial chitin
(Figure S3). In addition, an extra peak at 27.48 was observed in the lichen chitin that was not
present in the commercial chitin. The CrI value (83.8%) of the commercial chitin was higher
than that of the lichen chitin. As mentioned above, this indicates that there are b-1,3 glucan
residues in the lichen chitin.

2.5. Scanning electron microscopy

It was observed that the chitin of X. parietina had a smooth surface (Figure S4). Due to lichen
consisting of fungi and algae, the surface morphologies of the chitins obtained from fungi in
 4 M. Kaya et al.

earlier studies were considered. It was reported that the chitin samples obtained from some
fungal species were composed of nanofibres (Ifuku et al. 2011), while other chitin samples
obtained from different fungal species did not have nanofibres (Yen & Mau 2007). In this study,
the surface morphology of the chitin from X. parietina was smooth, as with other fungal chitins.

2.6. Elemental analysis

The elemental analysis results showed that the nitrogen (N), carbon (C) and hydrogen (H)
contents of the lichen chitin were 4.45%, 42.74% and 6.14%, respectively. Theoretically, the
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value of the N content in fully acetylated chitin is 6.89% (Ifuku et al. 2011). The value recorded
from the lichen chitin in this study was significantly lower than the theoretical value. Ifuku et al.
(2011) stated that the N content values for chitins obtained from different fungal species varied
between 2.96% and 6.37%. The N rate will be low in chitins obtained from some fungal species
because of the glucan residue (Ifuku et al. 2011). Considering that lichen consists of fungi, the
low N value is an expected result. Glucan residues were observed in the lichen chitin, as in
chitins obtained from other fungal species. The N values of the chitins extracted from insects,
such as cicada sloughs, bumble bee and Holotrichia parallela, were calculated to be 5.93%,
5.92% and 6.45%, respectively (Majtan et al. 2007; Sajomsang & Gonil 2010; Liu et al. 2012).
Even the N values of chitins obtained from organisms that are used to produce commercial chitin
varied between 4.85% and 6.24% (Majtan et al. 2007; Liu et al. 2012).

3. Conclusions
The lichen chitin is similar to fungal chitin due to it being in the a-form and containing glucan
residues, it is not similar to the chitins obtained from organisms such as crab, shrimp and krill
due to its smooth surface morphology. As already known, chitin and its derivatives are very
valuable biopolymers due to their multi-purpose uses. In this study, new and biotechnologically
important properties of X. parietina were revealed. This study suggests that X. parietina can be
used as an alternative source from which to obtain chitin. In the future, the obtained chitin can be
converted to chitosan and evaluated for use in areas related to chemistry, industry and medicine.

Supplementary material
The experimental part of this article is available online, alongside Table S1 and Figures S1– S5.

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