Original Article ISSN 0101-2061 (Print)

Food Science and Technology ISSN 1678-457X (Online)

DOI: https://doi.org/10.5327/fst.00287

Extraction and characterization of collagen from the skin

of Pterygoplichthys spp. (armored catfish) from Chiapas
Héctor Raziel LARA-JUACHE1 , Jorge Enrique WONG-PAZ2 , Xariss Miryam SÁNCHEZ-CHINO3 ,
Rebeca Isabel MARTÍNEZ-SALINAS4 , Everardo BARBA-MACÍAS5 , Juan Jesús MORALES-LÓPEZ1 ,
Anahí ARMAS-TIZAPANTZI1 , Arturo TORRES-DOSAL1*

Abstract
Because of its biocompatibility and safety, collagen is a valuable biomaterial that is used in various industries. While traditionally
sourced from terrestrial mammals, collagen extracted from fish waste is a promising alternative due to its chemical properties
and low cost. In Mexico, Pterygoplichthys spp. is considered an invasive species, inedible, and of limited commercial importance
due to the low muscle content and skin covered with rigid bony plates. This study aimed to extract and characterize collagen
from the skin of Pterygoplichthys spp. obtained from two communities in Chiapas, Mexico. Our results show that the extracted
collagen is type 1, with a yield of 43.0% (dry weight). Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis
revealed the presence of two alpha chains (α1, α2) and a beta and gamma component, consistent with type 1 collagen.
Liquid chromatography coupled to mass spectrophotometry analysis identified peptide sequences homologous to those
reported in other species. This study highlights the potential of Pterygoplichthys spp. skin collagen as a viable alternative to
mammalian collagen. The efficiency of the extraction process and the identification of peptides resembling those of the other
species underscore the feasibility of utilizing collagen from the skin of Pterygoplichthys spp. in industrial applications, offering
a sustainable solution to environmental and economic challenges. Practical Application: Collagen extracted from the skin of
Pterygoplichthys spp., an invasive fish species, can serve as a sustainable alternative to mammalian collagen, offering a cost-
effective biomaterial with properties suitable for various industrial applications.
Keywords: collagen extraction; Pterygoplichthys spp.; liquid chromatography coupled to mass spectrophotometry; sodium
dodecyl sulfate polyacrylamide gel electrophoretic profiling; type 1 collagen.

1 INTRODUCTION fish waste, including bones, scales, and skin, which has gained
attention for being a cheaper, higher yielding, and readily avail-
Collagen, a fundamental protein comprising approximately
able source (Chen et al., 2019; Hashim et al., 2015; Herath et al.,
30% of the total proteins in the cell matrix, plays a crucial role
2020; León-López et al., 2019; Zeng et al., 2009). Among the fish
in connective tissues such as blood vessels, corneas, bones, car-
tilage, tendons, and skin. It is widely used in various industries, species considered for collagen extraction, Pterygoplichthys spp.
including food, cosmetics, and drug formulation, particularly (armored catfish, devil fish, or plecos) stands out as a promising
in regenerative medicine and cell biology, due to its biocompat- source due to its abundance and low commercial value, attribut-
ibility, mechanical strength, and ability to support growth while ed to its low muscle content and skin covered with bony plates
preserving muscle rheological properties (Felician et al., 2018; (Ebenstein et al., 2015; Herath et al., 2020; Nurubhasha et al.,
Hashim et al., 2015; Herath et al., 2020; Nurubhasha et al., 2019). 2019). In Mexico, Pterygoplichthys spp. is considered an invasive
species that, due to its easy adaptability and wide distribution, has
Despite its versatility, the application of collagen is limited by caused socioeconomic and environmental problems (Amador-del
sociocultural practices as its primary source from mammals raises Ángel et al., 2016; Lorenzo-Márquez et al., 2016).
concerns regarding additional health costs and the transmission of
bovine and porcine diseases (Chen et al., 2019; Singh et al., 2011; Previous studies have highlighted the complete discarding
Yu et al., 2018). A potential solution lies in utilizing collagen from of Pterygoplichthys spp. by affected sectors, such as fishermen,

Received 29 Feb., 2024.

Accepted 16 Mar., 2024.
1Colegio de la Frontera Sur, San Cristóbal de las Casas, Chiapas, México.
2
Universidad Autónoma de San Luis Potosí, Facultad de Estudios Profesionales Zona Huasteca, San Luis Potosí, México.
3
CONAHCYT-Colegio de la Frontera Sur, Villahermosa, Tabasco, México.
4
Universidad de Ciencias y Artes de Chiapas, Facultad de Ingeniería Ambiental, Tuxtla Gutiérrez, Chiapas, México.
5
Colegio de la Frontera Sur, Villahermosa, Tabasco, México.
*Corresponding author: atorres@ecosur.mx
Conflict of Interest: nothing to declare.
Funding: Consejo Nacional de Ciencia, Humanidades y Tecnología, with Project number 319010, of the National Research and Incidence Projects on polluting processes,
toxic damage, and their socio-environmental impacts associated with sources of natural and anthropogenic origin; Project number 321911, of the scientific development
fund C-719/2022; graduate program grant (788816); and postdoctoral project (2668578).

Food Sci. Technol, Campinas, 44, e00287, 2024 1

 Extraction and characterization of collagen from the skin of Pterygoplichthys spp. (armored catfish) from Chiapas

in countries such as Mexico, India, and Indonesia, resulting in modifications according to our working conditions. We used
wasted protein-rich material (Ayala-Pérez et al., 2014; Herath 0.5 M acetic acid (CH3COOH) for 72 h at 4°C, with a mass/
et al., 2020; Nurubhasha et al., 2019). volume ratio of 1:20. At the end, the mixture was filtered through
a food grade mesh, and the pH was adjusted to 8.0 with 1 N
The use of this species in products of low commercial value,
NaOH. Precipitation was achieved by adding 2 M sodium chlo-
such as fertilizers and animal feed, has been studied to control
ride (NaCl) and incubating for 12 h at 4 °C, following the pro-
overpopulation caused by its invasion (Ayala-Pérez et al., 2015;
cedure described by Vidal et al. (2020). The precipitated sample
Prihanto et al., 2021). Some studies have also reported using the
was centrifuged at 6,000 rpm for 15 min at 4°C. The precipitate
skin for collagen extraction to generate value-added biomate-
was dialyzed through a 14 kDa molecular cutoff dialysis bag
rials (Ebenstein et al., 2015; Herath et al., 2020; Nurubhasha
(Sigma-Aldrich, D527-100FT) with 0.1 M CH3COOH for 48 h
et al., 2019). The use of the skin of Pterygoplichthys spp. could
at 4°C with agitation at 4,000 rpm and solution change every
be an attractive alternative to control the species’ invasion in the
12 h. This process was repeated with deionized water under the
aquatic systems of Chiapas, Mexico, reducing the impact that
same conditions as described above (Vidal et al., 2020; Yu et al.,
their presence has caused on biodiversity, such as silting of water
2018). Subsequently, the supernatant was lyophilized using a
bodies, erosion of riverbeds by their nests, turbidity generated
Labconco, Freezone 18 and stored at 4°C (Herath et al., 2020;
by the resuspension of sediments while they feed, pollution pro-
Singh et al., 2011; Zeng et al., 2012). The yield was calculated
duced by the decomposition of Pterygoplichthys spp. discarded
based on the ratio of the skin weight to the dry weight of the
by fishermen on the banks, as well as an increase in nutrients, sample (Zeng et al., 2012) (Equation 1).
especially nitrogen and phosphorous, originating from the ex-
crement of these fish in rivers and lagoons (Amador-del Ángel
et al., 2016; Ayala-Pérez et al., 2014; Capps & Flecker, 2013; Yield % = (Weight (g) of lyophilized
Herath et al., 2020; Lorenzo-Márquez et al., 2016). The aim of (1)
collagen/Weight (g) of dry skin used) x 100
this study was to extract and characterize collagen from the
skin of Pterygoplichthys spp., compare the methods used by
other authors, and generate a proposal for a strategy for further 2.3 Quantification of soluble protein
utilization in localities impacted by the fish’s precariousness. The soluble protein content in the collagen obtained from
the skin of Pterygoplichthys spp. was determined using the
2 MATERIALS AND METHODS Bradford method (Bradford, 1976) for which the sample was
prepared by dissolving 1 g of collagen in 0.5 M CH3COOH.
A total of 50 specimens of Pterygoplichthys spp. were Solutions of 100 μL of distilled water and collagen solution were
captured from the municipalities of La Flor de Chiapas prepared in a ratio of 1:10, 1:100, and 1:1,000 (v/v), then mixed
(17.487189379699288, -91.82607435206123) and Rómulo Calz- with 1,000 μL of Bradford reagent (Bio-Rad, Cat# 5000205)
ada (17.34261470132608, -93.55631205895111), Chiapas, Mex- composed of Coomassie Brilliant Blue G-250 dissolved in 55%
ico. The fish were sacrificed and preserved on ice for transport phosphoric acid and 15% methanol. After incubating for 5 min,
to the laboratory and refrigerated at 4°C. The skins were man- it was analyzed at 595 nm in a UV-visible spectrophotometer
ually removed, dried at 60°C for 24 h in a drying oven (Felisa, (Thermo Fisher, Genesys 150). Bovine serum gamma globulin
FE-133), then pulverized using a food processor (KitchenAid, at a concentration of 1 mg/mL (Bio-Rad, Cat# 5000208) was
KFP0919LCU), and sieved to a particle size of approximately used as a standard protein.
0.6 mm. The skin powder was stored at 4°C until further use
(Fonseka & Radampola, 2022).
2.4 Collagen characterization

2.1 Skin conditioning 2.4.1 Sodium dodecyl sulfate polyacrylamide gel electrophoretic
profiling (SDS-PAGE) and UV-visible spectroscopy
To remove noncollagen proteins, the powdered skin of
Pterygoplichthys spp. was treated with a 0.1 M sodium hydroxide To identify the type and purity of the collagen, SDS-PAGE
(NaOH) solution at a ratio of 1:10 (w/v) for 12 h at 4°C, with the was performed using a Mini-PROTEAN Tetra Cell system (Bio-
solution changed every 3 h. The wastewater was then washed Rad), according to the Laemmli method with slight modifications
with Milli Q ultrapure water until reaching a pH of 7.0 (Singh (Laemmli, 1970). The concentrator gel had a concentration of 4%,
et al., 2011; Zeng et al., 2012). Subsequently, the skin samples and a separator gel was prepared at 8%. The collagen sample was
were subjected to lipid removal using 10% butanol for 12 h at a solubilized in 0.1 M CH3COOH, mixed with 2 mL of Tris–HCl
ratio of 1:10 (w/v) at 4°C, followed by three washes in ultrapure sample buffer, and then heated at 95°C for 5 min. The mixture was
Milli Q water for 15 min (Herath et al., 2020; Singh et al., 2011; centrifuged at 5,000 rpm for 1 min. Each well was loaded with
Zeng et al., 2012). All the reagents used were of high-perfor- a volume of 10 μg/μL sample. The gel ran at 100 V for 60 min.
mance liquid chromatography grade. A type 1 collagen standard from rat tendon (Merck, Cat# 8-115)
was used for protein identification. Gels were stained with 0.1%
(w/v) Coomassie Brilliant Blue R-250 in 50% (v/v) methanol and
2.2 Extraction of collagen in acetic acid
6.8% (v/v) glacial acetic acid for 5 h, then destained using 7.5%
Collagen extraction was performed according to the meth- (v/v) glacial acetic acid and 5% (v/v) methanol (El-Rashidy et al.,
odology described by Nurubhasha et al. (2019), with some 2015; Nurubhasha et al., 2019; Vidal et al., 2020).

2 Food Sci. Technol, Campinas, 44, e00287, 2024

 LARA-JUACHE et al.

The UV-visible absorption spectrum was determined using density, which can alter electrostatic interactions and protein
a spectrophotometer (Thermo Fisher, Genesys 150) in the wave- structure (Tan & Chang, 2018). Moreover, the solvent and raw
length range of 190–400 nm at a scan rate of 1 nm/s. The collagen material determine the yield by the effective collision between
sample was solubilized in 0.5 M CH3COOH at a 1:1 (m/v) ratio both components; this interaction influences the matter transfer
(Nazeer et al., 2014; Nurubhasha et al., 2019; Sampath Kumar processes. This reflects a higher or lower yield in the collagen
& Nazeer, 2013). extraction as determined by Lewis’ theory of coalitions estab-
lished in 1918. There is a critical concentration depending on
2.4.2 Amino acid sequencing in collagen the solvent/raw material ratio; increasing the ratio between the
components will no longer benefit the process and the yield, as
The amino acid sequence of collagen was determined well as increasing the extraction time (Chen et al., 2016; Tan
according to the method described by Chinh et al. (2019). & Chang, 2018). This is due to the mechanism of action of the
For this purpose, after the SDS-PAGE process, the bands of organic acid during protein extraction. The direct interaction
interest obtained in the polyacrylamide gel were cut, washed, of acetic acid with the hydrogen bridges present in the collagen
and analyzed by liquid chromatography coupled to mass triple helix separates the bonds between the alpha helix chains,
spectrophotometry (LC-MS/MS). The results obtained were causing the protein to solubilize in the medium (Tan & Chang,
searched and compared in the protein database of the National 2018; Zeng et al., 2012). This mechanism is affected if the solvent/
Center for Biotechnology Information. raw material concentration is inadequate.

3 RESULTS AND DISCUSSION 3.2 Quantification and identification of protein in collagen

3.1 Collagen extraction The concentration of soluble protein in the collagen ex-
tracted from Pterygoplichthys spp. was 69.7 mg/mL, higher
We obtained a yield of 43.05 ± 0.05% of acetic acid soluble than that obtained by Nurubhasha et al. (2019), who reported a
collagen (ASC) from the skin of Pterygoplichthys spp. on a dry concentration of 22.56 mg/mL. The difference in concentration
weight basis (freeze-dried). The yield obtained under our oper- could be due to the experimental conditions such as collagen ex-
ating conditions was higher than that reported by Herath et al. traction time and raw material/solvent concentration similar to
(2020), who obtained a yield of 26.20% from Pterygoplichthys the dialysis process. The increase in protein concentration may
spp. skin caught in Lake Tempe, South Sulawesi, Indonesia. be due to a lower loss of protein when dialysis was performed to
Nurubhasha et al. (2019) reported a lower yield of 19.6% on remove NaCl ions for collagen precipitation. Figure 1A shows
wet weight basis from the skin of Pterygoplichthys spp. obtained the electrophoretic pattern determined by protein mobility
from Kolleru Lake, Andhra Pradesh. in SDS-PAGE. Line 1 corresponds to the molecular weight
The difference in collagen extraction yield can be attributed marker, line 2 to the type 1 collagen standard, and line 3 to the
to the slight modifications in the methodology used. For instance, collagen extract obtained. The modeled gel shows alpha 1 (α1)
our study employed a 12-h period for globular protein removal and alpha 2 (α2) chains, weighing approximately 130 and 120
using a 0.1 M NaOH concentration and pH 11. In contrast, kDa, respectively, which form the collagen triple helix. Also,
Herath et al. (2020) and Nurubhasha et al. (2019) conducted the high-molecular-weight components were observed, called β
extractions with an incubation period of 4 and 36 h using NaOH chain with a molecular weight of 250 kDa, formed by two α1
concentrations of 0.5 and 0.1 M, respectively. Furthermore, the chains in laminar form crosslinked by hydrogen bridges and γ
collagen extraction time was 24 h (Nurubhasha et al., 2019); on chain (gamma) with a molecular weight higher than 250 kDa,
the contrary, Herath et al. (2020) used a period of 72 h, like the formed by three chains α1 (Chen et al., 2016). Therefore, the
one implemented in this study. However, it should be noted that ASC extracted in this study was classified as type 1 collagen.
the raw material/solvent ratio for this study was 1:20, whereas These results are similar to those reported by Nurubhasha et al.
the aforementioned authors used a ratio of 1:10. Likewise, the (2019), who extracted type 1 collagen from acetic acid Pterygo-
difference in collagen yield could also be attributed to the un- plichthys spp. skin. Furthermore, Singh et al. (2011) and Tan and
Chang (2018) reported the extraction of type 1 collagen from the
equal cross-linking of the collagen molecule. Highly cross-linked
skin of different Pterygoplichthys spp. species and report bands
nonhelical terminal ends of the telopeptide region can decrease
with molecular weights of 100–130, 90–120, and 200–260 kDa
collagen solubility in the acid solvent (Abinaya & Gayathri, 2019;
corresponding to α1, α2, β, and γ chains, respectively.
Zeng et al., 2012). Previous studies have shown that collagen is
soluble in 0.5 M acetic acid (Abinaya & Gayathri, 2019; Kuwa- The UV-visible characterization was analogous to that re-
hara, 2021; Zeng et al., 2012). However, slight modifications in ported by Nurubhasha et al. (2019), confirming that the pro-
the experimental and preparative conditions of the methodology tein obtained is type 1 collagen, as well as coinciding with the
play a key role in the extraction performance of collagen, and electrophoresis analysis. Figure 1B shows the absorption spec-
factors such as pH, solvent concentrations (sodium hydrox- trum in a spectral sweep from 190 to 400 nm of the collagen
ide and acetic acid), particle size of the sample, weight/volume of the Pterygoplichthys spp. and the type 1 collagen standard.
ratio, and interaction time (raw materials/solvent) influence The similarity of absorbance in the sweep can be seen, with
the extraction process (Chen et al., 2016; Nurubhasha et al., the maximum absorption length found at 236 and 235 nm,
2019; Tan & Chang, 2018; Zeng et al., 2012). For example, pH respectively. This absorption is mainly attributed to the n–n*
can reduce collagen solubility by affecting the protein’s charge bonds of the C = O, -COOH, and CONH2 groups present in

Food Sci. Technol, Campinas, 44, e00287, 2024 3

 Extraction and characterization of collagen from the skin of Pterygoplichthys spp. (armored catfish) from Chiapas

the polypeptide chain of collagen (Abinaya & Gayathri, 2019; (Zeng et al., 2012). However, the collagen sample obtained in
Chen et al., 2019). The ultraviolet region of proteins generates this study showed a low absorbance at this length as shown
absorbance near 280 nm; this signal corresponds to aromatic in Figure 1B. The inability to absorb at higher UV regions is
amino acids such as tyrosine, phenylalanine, and tryptophan related to tyrosine, phenylalanine, and tryptophan deficiency

Figure 1. SDS-PAGE and UV-visible spectrophotometric analysis of ASC obtained from Pterygoplichthys spp. skin. (A) Line 1: molecular weight
marker. Line 2: collagen type 1 standard (STD). Line 3: extracts from ASC. (B) UV-visible spectrum. Continuous red line: acid-soluble collagen
from Pterygoplichthys spp. Dotted black line: collagen type 1 standard.

Figure 2. Sequencing of collagen peptides. (A) Chromatogram of total ions released from the α1 chain of Pterygoplichthys spp. after 18 h of
enzymatic digestion. (B) MS/MS spectrum corresponding to ion 739.36. (C) Chromatogram of total ions released from the α2 chain. (D) MS/
MS spectrum corresponding to the 747.87 ion released from the α2 chain.

4 Food Sci. Technol, Campinas, 44, e00287, 2024

 LARA-JUACHE et al.

in Pterygoplichthys spp. collagen (Abinaya & Gayathri, 2019; in SDS-PAGE (Figure 1A) was analyzed using the same method
Nurubhasha et al., 2019; Zeng et al., 2012). Nurubhasha et al. as above; according to the mass spectrum (Figure 2D), the
(2019) reported analogous findings, obtaining collagen from the peptide GPSGSVGGPAGPAGPAGAR was identified with an
same species with a maximum absorbance at 235 nm. m/z of 747. 8, a molecular weight of 1,542.80 Da, and a charge
of +2, which was found in the α2 chain of type 1 collagen from
I. punctatus (Table 1), and its retention peak was identified at
3.3 Collagen identification (amino acid sequencing)
minute 41.37 (Figure 2C).
Peptide sequence identification by LC-MS/MS of collagen
SDS-PAGE was used to analyze the composition and molec-
is shown in Figure 2. Figure 2A shows the total ion chro-
ular weight distribution of the collagen obtained. The enzymatic
matogram of the peptide mixture released after the enzymatic digestion of the chains identified in SDS-PAGE (Figure 1A)
digestion performed on the identified α1 band in SDS-PAGE was performed, followed by analysis with LC-MS/MS for the
(Figure 1A); this peptide mixture was useful to corroborate the amino acid sequences of collagen. Additionally, BLAST analysis
type of collagen obtained (Kleinnijenhuis et al., 2022; Zhang was used to study the homology of the sequences, where the
et al., 2006). The mass spectrum was implemented to determine number of coincident peptides and the coverage of the collagen
the m/z of the ions and their charge states (Figure 2B), where chain sequences were evaluated as the main parameters for their
the ion with m/z 739.36 retained at minute 30.29 in the ion identification (Chen et al., 2017; Chinh et al., 2019; Huang et al.,
chromatogram was manually identified to avoid assignment 2015; Zeng et al., 2012).
errors as GPSGPAGAAGPAGPR (Kleinnijenhuis et al., 2022;
Richter et al., 2020). A molecular weight of 1,477.72 Da was Two matching peptides were identified from the α1 chain of
manually calculated for this peptide. The ion had a charge of +2, type 1 collagen reported in species such as I. punctatus, Ictalurus
indicating that the peptide is charged differently (Chen et al., furcatus, Oreochromis niloticus, and Cyprinus carpio (Table 1),
2018), that is, there are peaks with m/z above 739.36 as seen in in which their coverage in the α1 chain sequence was 13% for
the MS/MS spectrum (Figure 2B), and the 921.48 peak was the each species, similar to that reported by Zeng et al. (2012). Fur-
most abundant peak generated from the cleavage of the Pro-Gly thermore, variations between peptide positions were found to
peptide bond. Both peaks 753.40 and 978.50 were of relatively be given by only one unit, that is, the GPAGPAGAAGPAGPR
high abundance and formed from the cleavage of Gly-Ala and peptide was found at position 1,056–1,070 for I. punctatus and
Gly-Ser peptide bonds, respectively. The peak at m/z 1162.29 at position 1,054–1,068 for O. niloticus.
generated by cleavage of the Pro-Gly peptide bond had a rel- Harvey et al. (2018) mentioned that the ion found in MS/
atively low abundance. The peptide (GPSGPAGAAGPAGPR) MS is known as a collagen precursor ion because the peptide
was found in the α1 chain of type 1 collagen of Ictalurus punc- repeats, that is, it can be found in the collagen of other fish spe-
tatus (Table 1). The digested mixture of the α2 chain identified cies. However, Kleinnijenhuis et al. (2022) mentioned that the

Table 1. Summary of LC-MS/MS results based on those obtained by SDS-PAGE.

Experimental protein Protein description No. adhesion Mass (Da) Score Number of peptides Matched amino acid sequence

Collagen type 1 alpha 1 604 GAPGAPGPA 612

XP_017348849.2 137,314 3,487 2
(Ictalurus punctatus) 1057 GPSGPAGAAGPAGPR 1071
Collagen, type 1, alpha 1 603 GAPGAPGPA 611
XP_053469073.1 137,260 3,478 2
(Ictalurus furcatus) 1056 GPSGPAGAAGPAGPR 1070
ASC alpha 1
Collagen type 1 alpha 1054 GPSGPAGAAGPAGPR 1068
NP_001266373.1 134,883 3,444 2
1 (Oreochromis niloticus) 1027 GAPGAPGP 1034
Collagen, type 1, alpha 1 602 GAPGAPGPA 610
XP_042576683.1 136,931 2,825 2
(Cyprinus carpio) 1055 GPSGPAGAAGPAGPR 1069
479 PGNIGFPGPK 488
Collagen type 1, alpha 2 972 GPSGSVGPAGPAGAR 986
XP_047006669.2 126,437 3,757 2
(Ictalurus punctatus) 853 GPAGPPGA 860
305 PGVAGTPGF 313
479 PGNIGFPGPK 488
Collagen type 1, alpha 2 972 GPSGSVGPAGPAGAR 986
XP_053468726.1 126,608 3,742 2
(Ictalurus furcatus) 853 GPAGPPGA 860
305 PGVAGTPGF 313
ASC alpha 2
478 GNIGFPGPK 486
Collagen type 1, alpha 877 G-VGEPGR 884
NP_001269826.1 124,065 2,137 4
2 (Oreochromis niloticus) 886 VGPAGPPGA 894
770 PPGLTGFP 777
479 PGNIGFPGPK 488
Collagen type 1, alpha 2 879 G-VGEPGR 886
XP_042601316.1 127,304 1,967 4
(Cyprinus carpio) 815 VGPAGPPG 822
772 PPGLTGFP 779

Food Sci. Technol, Campinas, 44, e00287, 2024 5

 Extraction and characterization of collagen from the skin of Pterygoplichthys spp. (armored catfish) from Chiapas

content and variation of amino acids in the collagen sequence In A. M. Low Pfeng, P. A. Quijón & E. M. P. Recagno (eds.). Especies
is different between one species and another; therefore, in other invasoras acuáticas: casos de estudio en ecosistemas de México (p.
sequences, the change in amino acid could be found, for exam- 313-336). Secretaría de Medio Ambiente y Recursos Naturales.
ple, GSAGPAGAGAAGPAGPR and GPAGPAGAAGPAGPR, Ayala-Pérez, L. A., Vega-Rodríguez, B. I., & González-Greicy, J. (2015).
where S is the change (Huang et al., 2015). Furthermore, four El pez diablo en México guía para administradores y usuarios de
peptides of the α2 chain of collagen type 1 reported in the spe- recursos pesqueros. México Universidad Autónoma Metropolitana.
cies I. punctatus, I. furcatus and four peptides for the species O. Bradford, M. M. (1976). A rapid and sensitive method for the quanti-
niloticus and C. carpio were identified (Table 1); these results are tation of microgram quantities of protein utilizing the principle
superior to those reported by Chen et al. (2017) and Zeng et al. of protein-dye binding. Analytical Biochemistry, 72(1-2), 248-254.
(2012). The sequence coverage of the collagen chain was 6%; https://doi.org/10.1006/abio.1976.9999
it was found that the PGNIGFPGPK peptide has a variation of Capps, K. A., & Flecker, A. S. (2013). Invasive fishes generate biogeo-
one unit between I. punctatus and C. carpio, while for the other chemical hotspots in a nutrient-limited system. PLoS One, 8(1),
peptides, the position variations were more than one unit (Chen e54093. https://doi.org/10.1371%2Fjournal.pone.0054093
et al., 2018; Huang et al., 2015; Kleinnijenhuis et al., 2022). Chen, J., Li, M., Yi, R., Bai, K., Wang, G., Tan, R., Sun, S., & Xu, N.
(2019). Electrodialysis extraction of pufferfish skin (Takifugu
The results presented above revealed that the extracted col- flavidus): A promising source of collagen. Marine Drugs, 17(1),
lagen possessed a higher similarity to the species mentioned 25. https://doi.org/10.3390%2Fmd17010025
above in relation to the α1 and α2 chains of type 1 collagen. Chen, J., Li, L., Yi, R., Gao, R., & He, J. (2018). Release kinetics of Tilapia
The relative molecular weights (α1: 136,656 Da; α2: 126,472 Da) scale collagen I peptides during tryptic hydrolysis. Food Hydrocol-
are analogous to the SDS-PAGE results (α1: 130,000 Da; α2: loids, 77, 931-936. https://doi.org/10.1016/j.foodhyd.2017.11.040
120,000 Da), indicating that the collagen extracted corresponds Chen, J., Li, L., Yi, R., Xu, N., Gao, R., & Hong, B. (2016). Extraction
to a type 1 collagen (Chen et al., 2018). Furthermore, based on the and characterization of acid-soluble collagen from scales and
percentage of protein sequence coverage, it can be confirmed that skin of tilapia (Oreochromis niloticus). LWT-Food Science and
the amino acid sequence of the collagen extracted from the skin Technology, 66, 453-459. https://doi.org/10.1016/j.lwt.2015.10.070
of Pterygoplichthys spp. was very similar to that of I. punctatus. Chen, J., Liu, Y., Yi, R., Li, L., Gao, R., Xu, N., & Zheng, M. (2017). Charac-
terization of collagen enzymatic hydrolysates derived from lizardfish
(Synodus fuscus) scales. Journal of Aquatic Food Product Technology,
4 CONCLUSION
26(1), 86-94. https://doi.org/10.1080/10498850.2015.1094687
The results showed that the extraction process in 0.5 M acetic Chinh, N. T., Manh, V. Q., Trung, V. Q., Lam, T. D., Huynh, M. D.,
acid for 72 h at pH 2.5 implemented the skin of Pterygoplichthys Tung, N. Q., Trinh, N. D., & Hoang, T. (2019). Characterization
spp. with a 0.1 M sodium hydroxide pretreatment for 6 h at pH of collagen derived from tropical freshwater carp fish scale wastes
11.0 that allowed us to obtain a recovery yield of 43.05% of colla- and its amino acid sequence. Natural Product Communications,
gen. The collagen extracted from Pterygoplichthys spp. was charac- 14(7). https://doi.org/10.1177/1934578X19866288
terized chemically and identified as type 1 collagen, confirmed by Ebenstein, D., Calderon, C., Troncoso, O. P., & Torres, F. G. (2015).
SDS-PAGE, UV-visible, and LC-MS/MS techniques. The presence Characterization of dermal plates from armored catfish Pterygo-
of two alpha chains (α1 and α2) and β and γ components supports plichthys pardalis reveals sandwich-like nanocomposite structure.
the typical structure of type 1 collagen. Pterygoplichthys spp. skin Journal of the Mechanical Behavior of Biomedical Materials, 45,
is a promising source of type 1 collagen, being a viable alternative 175-182. https://doi.org/10.1016/j.jmbbm.2015.02.002
for the use of mammalian-derived collagen used in the food, El-Rashidy, A. A., Gad, A., Abu-Hussein, A. E.-H. G., Habib, S. I.,
cosmetic, and biomedical industries. It is important to point out Badr, N. A., & Hashem, A. A. (2015). Chemical and biological
that future studies are required to assess its complete effectiveness evaluation of Egyptian Nile Tilapia (Oreochromis niloticas) fish
scale collagen. International Journal of Biological Macromolecules,
in terms of extraction since this work constitutes a preliminary
79, 618-626. https://doi.org/10.1016/j.ijbiomac.2015.05.019
alternative for the exploitation of a fish that generates environ-
mental and economic problems in the areas where it is present. Felician, F. F., Xia, C., Qi, W., & Xu, H. (2018). Collagen from marine
biological sources and medical applications. Chemistry & Biodi-
versity, 15(5), e1700557. https://doi.org/10.1002/cbdv.201700557
REFERENCES Fonseka, R., & Radampola, K. (2022). Feasibility of using sailfin cat-
Abinaya, M., & Gayathri, M. (2019). Biodegradable collagen from fish meal as an alternative to commercial fishmeal in the diets of
Scomberomorus lineolatus skin for wound healing dressings and juvenile guppy (Poecilia reticulata). Journal of Fisheries, 10(1),
its application on antibiofilm properties. Journal of Cellular Bio- 101203. https://doi.org/10.17017/j.fish.344
chemistry, 120(9), 15572-15584. https://doi.org/10.1002/jcb.28824 Harvey, V. L., Daugnora, L., & Buckley, M. (2018). Species identification
Amador-del Ángel, L., Wakida-Kusunoki, A., Sánchez-Martínez, M., & of ancient Lithuanian fish remains using collagen fingerprint-
Hernández-Nava, J. (2016). Consideraciones económicas para el ing. Journal of Archaeological Science, 98, 102-111. https://doi.
manejo del pez diablo en el área protegida de Laguna de Términos. org/10.1016/j.jas.2018.07.006
In L. A. Ayala-Pérez, B. I. Vega-Rodríguez & J. González-Greicy Hashim, P., Ridzwan, M. M., Bakar, J., & Hashim, M. D. (2015). Colla-
(Ed.). El pez diablo en México: Protocolo de prevención, respuesta gen in food and beverage industries. International Food Research
rápida y control (p. 144-156). Universidad Autónoma Metropolitana. Journal, 22(1), 1-8.
Ayala-Pérez, L. A., Pineda-Peralta, A. D., Álvarez-Guillen, H., & Ama- Herath, H., Kalutharage, N. K., & Cumaranatunga, P. R. T. (2020). Solutions
dor-del Ángel, L. E. (2014). El pez diablo (Pterygoplichthys spp.) to an alien species invasion from aquarium aquaculture: Isolation
en las cabeceras estuarinas de la Laguna de Términos, Campeche. and characterization of acid soluble collagen from sailfin catfish,

6 Food Sci. Technol, Campinas, 44, e00287, 2024

 LARA-JUACHE et al.

Pterygoplichthys disjuctivus (Weber, 1991) in Sri Lanka. Sri Lanka Richter, K. K., McGrath, K., Masson-MacLean, E., Hickinbotham, S.,
Journal of Aquatic Sciences, 25(1), 19-32. https://doi.org/10.4038/ Tedder, A., Britton, K., Bottomley, Z., Dobney, K., Hulme-Beaman,
sljas.v25i1.7573 A., & Zona, M. (2020). What’s the catch? Archaeological appli-
Huang, B.-B., Lin, H.-C., & Chang, Y.-W. (2015). Analysis of proteins cation of rapid collagen-based species identification for Pacific
and potential bioactive peptides from tilapia (Oreochromis spp.) Salmon. Journal of Archaeological Science, 116, 105116. https://
processing co-products using proteomic techniques coupled with doi.org/10.1016/j.jas.2020.105116
BIOPEP database. Journal of Functional Foods, 19(part A), 629- Sampath Kumar, N., & Nazeer, R. (2013). Characterization of acid
640. https://doi.org/10.1016/j.jff.2015.09.065 and pepsin soluble collagen from the skin of horse mackerels
Kleinnijenhuis, A. J., van Holthoon, F. L., & van der Steen, B. (2022). (Magalaspis cordyla) and croaker (Otolithes ruber). International
Identification of collagen 1α3 in teleost fish species and typical Journal of Food Properties, 16(3), 613-621. https://doi.org/10.108
collision induced internal fragmentations. Food Chemistry: X, 14, 0/10942912.2011.557796
100333. https://doi.org/10.1016/j.fochx.2022.100333 Singh, P., Benjakul, S., Maqsood, S., & Kishimura, H. (2011). Iso-
Kuwahara, J. (2021). Extraction of type I collagen from tilapia scales lation and characterisation of collagen extracted from the
using acetic acid and ultrafine bubbles. Processes, 9(2), 288. https:// skin of striped catfish (Pangasianodon hypophthalmus).
doi.org/10.3390/pr9020288 Food Chemistry, 124(1), 97-105. https://doi.org/10.1016/j.
Laemmli, U. K. (1970). Cleavage of structural proteins during the foodchem.2010.05.111
assembly of the head of bacteriophage T4. Nature, 227(5259), Tan, Y., & Chang, S. K. (2018). Isolation and characterization of
680-685. https://doi.org/10.1038/227680a0 collagen extracted from channel catfish (Ictalurus punctatus)
León-López, A., Morales-Peñaloza, A., Martínez-Juárez, V. M., Var- skin. Food Chemistry, 242, 147-155. https://doi.org/10.1016/j.
gas-Torres, A., Zeugolis, D. I., & Aguirre-Álvarez, G. (2019). foodchem.2017.09.013
Hydrolyzed collagen—Sources and applications. Molecules, 24(22), Vidal, A. R., Duarte, L. P., Schmidt, M. M., Cansian, R. L., Fernandes,
4031. https://doi.org/10.3390%2Fmolecules24224031 I. A., de Oliveira Mello, R., Demiate, I. M., & Dornelles, R. C. P.
Lorenzo-Márquez, H., Torres-Dosal, A., Barba Macías, E., Ilizaliturri (2020). Extraction and characterization of collagen from sheep
Hernández, C. A., Martínez-Salinas, R. I., Morales López, J. J., & slaughter by-products. Waste Management, 102, 838-846. https://
Sánchez Moreno, I. (2016). Estimación de riesgo de exposición a doi.org/10.1016/j.wasman.2019.12.004
metales pesados por consumo de plecos (Pterygoplichthys spp.) en Yu, F., Zong, C., Jin, S., Zheng, J., Chen, N., Huang, J., Chen, Y., Huang,
infantes de comunidades ribereñas de los ríos Grijalva y Usumac- F., Yang, Z., & Tang, Y. (2018). Optimization of extraction con-
inta, México. Revista Internacional de Contaminación Ambiental, ditions and characterization of pepsin-solubilised collagen from
32(2), 153-164. https://doi.org/10.20937/RICA.2016.32.02.02 skin of giant croaker (Nibea japonica). Marine Drugs, 16(1), 29.
Nazeer, R., Kavitha, R., Ganesh, R. J., Naqash, S. Y., Kumar, N. S., & https://doi.org/10.3390/md16010029
Ranjith, R. (2014). Detection of collagen through FTIR and HPLC Zeng, S., Yin, J., Yang, S., Zhang, C., Yang, P., & Wu, W. (2012).
from the body and foot of Donax cuneatus Linnaeus, 1758. Jour- Structure and characteristics of acid and pepsin-solubilized
nal of Food Science and Technology, 51(4), 750-755. https://doi. collagens from the skin of cobia (Rachycentron canadum).
org/10.1007/s13197-011-0539-1 Food Chemistry, 135(3), 1975-1984. https://doi.org/10.1016/j.
Nurubhasha, R., Sampath Kumar, N., Thirumalasetti, S. K., Simhacha- foodchem.2012.06.086
lam, G., & Dirisala, V. R. (2019). Extraction and characterization of Zeng, S., Zhang, C., Lin, H., Yang, P., Hong, P., & Jiang, Z. (2009).
collagen from the skin of Pterygoplichthys pardalis and its poten- Isolation and characterisation of acid-solubilised collagen from
tial application in food industries. Food Science and Biotechnology, the skin of Nile tilapia (Oreochromis niloticus). Food Chemistry,
28(6), 1811-1817. https://doi.org/10.1007/s10068-019-00601-z 116(4), 879-883. https://doi.org/10.1016/j.foodchem.2009.03.038
Prihanto, A. A., Nurdiani, R., & Sari, L. W. (2021). Production and Zhang, G., Sun, A., Li, W., Liu, T., & Su, Z. (2006). Mass spectrometric
characteristics of sailfin catfish (Pterygoplichthys pardalis) protein analysis of enzymatic digestion of denatured collagen for identi-
hydrolysate. F1000Research, 10, 1089. https://doi.org/10.12688/ fication of collagen type. Journal of Chromatography A, 1114(2),
f1000research.73335.1 274-277. https://doi.org/10.1016/j.chroma.2006.03.039

Food Sci. Technol, Campinas, 44, e00287, 2024 7