Current Research in Biotechnology 4 (2022) 119–128

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Current Research in Biotechnology

journal homepage: www.elsevier.com/locate/crbiot

Influence of gamma irradiation on the properties of bacterial cellulose

produced with concord grape and red cabbage extracts
Helenise Almeida do Nascimento a, Julia Didier Pedrosa Amorim b,c, Cláudio José Galdino da Silva Júnior b,c,
Alexandre D'Lamare Maia de Medeiros b,c, Andréa Fernanda de Santana Costa c,d, Daniella Carla Napoleão a,
Glória Maria Vinhas a, Leonie Asfora Sarubbo c,e,⇑
a
Centro de Tecnologia e Geociências, Universidade Federal de Pernambuco (UFPE), Cidade Universitária, s/n, CEP: 50740-540, Recife, Brazil
b
Rede Nordeste de Biotecnologia (RENORBIO), Universidade Federal Rural de Pernambuco (UFRPE), Rua Dom Manuel de Medeiros, s/n – Dois Irmãos, CEP: 52171-900, Recife, Pernambuco,
Brazil
c
Instituto Avançado de Tecnologia e Inovação (IATI), Rua Potyra, n. 31, Prado, CEP: 50751-310, Recife, Pernambuco, Brazil
d
Centro de Comunicação e Design, Centro Acadêmico da Região Agreste, Universidade Federal de Pernambuco (UFPE), BR 104, Km 59, s / n – Nova Caruaru, Caruaru, Pernambuco CEP:
50670-90, Brazil
e
Escola UNICAP Icam Tech, Universidade Católica de Pernambuco (UNICAP), Rua do Príncipe, n. 526, Boa Vista, CEP: 50050-900, Recife, Pernambuco, Brazil

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

Bacterial cellulose (BC) is a biopolymer produced by microorganisms that has high purity, non‐toxicity,
biodegradability, and biocompatibility. These characteristics make BC suitable for applications in sectors such
as the cosmetic, biomedical, and packaging industries. To ensure biosafety for use in these fields, an efficient
sterilisation process is necessary. Gamma irradiation is often used as a sterilisation technique, but it has high
penetration power. Therefore, evaluating its effects on the physicochemical properties of BC after exposure is
important. To date, few studies have reported BC produced with alternative media and sterilised with 60Co
radiolytic irradiation. In this work, the bacterium Komagataeibacter hansenii (formerly known as
Gluconacetobacter hansenii) was used for the production of BC in standard Hestrin‐Schramm medium and alter-
native media formulated with different compositions of concord grape and red cabbage extracts. The samples
were sterilised with 60Co irradiation. Comparisons of the samples before and after exposure to gamma irradi-
ation were performed with X‐ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), thermo-
gravimetric analysis, and scanning electron microscopy (SEM). The results revealed no substantial changes
in the membrane characteristics after irradiation with a dose of 25 kGy. SEM and FTIR analyses demonstrated
the preservation of the polymeric structure of the BC. XRD demonstrated that the process did not create new
planes of symmetry. With the exception of one sample, the mean reduction in crystallinity was 9.96%. The pre-
sent findings show that the polymer is safe and has potential for use in the cosmetic, biomedical, and food
industries.

Introduction gataeibacter (formerly known as Gluconacetobacter), Rhizobium,

The synthesis of bacterial cellulose (BC) was first reported by
Adrian J. Brown in 1886. Bacteria able to synthesize cellulose can
be considered ubiquitous since they have been found to inhabit differ-
ent ecosystems. They can be isolated from environmental sources
(such as soil and plants), from insects, humans, and from food sources
(La China et al., 2021). During an acetic fermentation process, the for-
mation of a gelatinous film was observed on the surface of a liquid cul-
ture medium (Brown, 1886; Parte et al., 2020). This film has been
reported to be synthesised by certain genera of bacteria, such as Koma-
Agrobacterium, Rhodobacter, and Sarcina (Andree et al., 2021;
Azeredo et al., 2019; Yamada et al., 2012). The genus Komagataeibacter
includes species, such as K. xylinus, K. europaeus and K. hansenii, that
have been described as the highest cellulose producing organisms
(Gullo et al., 2018). An important reservoir in which it is also possible
to detect strains belonging to the Komagataeibacter genus is kombucha
tea, a fermented beverage traditionally produced in Asia. Kombucha
tea is characterized by a microbial community in which different
microbial groups, mainly yeasts and bacteria, live in a symbiotic life-
style (Arıkan et al., 2020; La China et al., 2021).

⇑ Corresponding author at: Escola Icam Tech, Universidade Católica de Pernambuco (UNICAP), Rua do Príncipe, n. 526, Boa Vista, CEP: 50050-900, Recife, Pernambuco, Brazil.
E-mail address: leonie.sarubbo@unicap.br (L. Asfora Sarubbo).

https://doi.org/10.1016/j.crbiot.2022.02.001
Received 26 October 2021; Revised 8 February 2022; Accepted 9 February 2022

2590-2628/© 2022 The Authors. Published by Elsevier B.V.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
H. Almeida do Nascimento et al.
BC is an extracellular metabolic product with a fundamental phys-
iological role in the lifecycle of the microorganism, serving as a protec-
tive barrier and increasing in oxygen availability and the transport of
nutrients for the enhancement of microbial growth (Gromovykh et al.,
2017).
Studies have shown that the biopolymer has a high degree of pur-
ity, non‐toxicity, high porosity, high water holding capacity, and a
nanofibrillar structure (Gallegos et al., 2016; Inoue et al., 2020). This
combination of characteristics makes BC a highly versatile polymer for
applications in various fields (Phruksaphithak et al., 2019).
Several factors must be considered during the BC production pro-
cess to determine the resulting set of morphological characteristics,
physical properties, and production yield. Research has indicated that
factors such as pH, temperature, oxygen regulation, bacterial strain,
fermentation time, inoculation volume, and composition of the culture
medium influence the yield and final properties of BC (Wang et al.,
2019; Liu et al., 2017; Campano et al., 2015; Tang et al., 2010).
Hestrin‐Schramm (HS) medium (Hestrin and Schramm, 1954) is
traditionally used for BC production but it is expensive because it
requires a high concentration of nutrients, thereby limiting its produc-
tion (Costa et al., 2017; Blanco et al., 2018). Thus, attempts have been
made to find alternative nutrient sources to improve the benefit‐cost
ratio, achieve higher yields, and enable industrial applications on lar-
ger scales (Amorim et al., 2019; Revin et al., 2018). The use of substi-
tutes can be fully in natura (Machado et al., 2018) or can consist of
partial replacement of certain compounds of the medium (Amorim
et al., 2019), with the presence of synthetic compounds and supple-
mentation with other nutrients.
In applications in the biomedical, pharmaceutical, cosmetic, and
food packaging industries, the use of biopolymers requires specific
sterilisation methods to inactivate potentially harmful microorganisms
and avoid serious human infections (Beh et al., 2019). However, the
traditional BC purification method using NaOH at a temperature of
90 °C [as described by Hwang et al. (1999)] may not be capable of
inactivating all bacterial cells entrapped in the nanofibers of the BC,
thus resulting in bacterial growth when the membrane is inoculated
again in the production medium (Amorim et al., 2021). Depending
on the application of the polymer, this factor can be worrisome.
Gamma irradiation, an ionic process without heat, is often used for
the sterilisation of pharmaceutical products and biomedical devices
(Galante et al., 2018) as well as the preservation of foods (Jipa
et al., 2012), with doses ranging from 25 (most common) to 40 kGy
(Hodder et al., 2019; Paula et al., 2019).
This simple, effective procedure with high penetration power (Jipa
et al., 2012) is also widely used for polymer sterilisation (Paula et al.,
2019; Benyathiar et al., 2021). When misused, however, gamma irra-
diation can cause crosslinking or degradation of the polymer chain
(Rogers, 2012), thereby altering various physical and chemical proper-
ties of BC (Salari et al., 2021; Paula et al., 2019; Hodder et al., 2019).
The objective and innovation of this work was to evaluate the
effects of gamma irradiation on the physical and chemical properties
of BC through experimental characterisation techniques, such as Four-
ier transform infrared spectroscopy (FTIR), X‐ray diffraction (XRD),
thermogravimetric analysis (TGA), and scanning electron microscopy
(SEM).
Materials and methods
Materials
Glucose, peptone, yeast extract, citric acid monohydrate, agar, dis-
odium phosphate, and sodium hydroxide were purchased from Merck
Ltd., USA. Concord grape (Vitis labrusca) and red cabbage (Brassica
oleracea var. capitata f. rubra) were obtained from a local market in
the city of Recife, Brazil.
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Current Research in Biotechnology 4 (2022) 119–128
Microorganism
The microorganism Komagataeibacter hansenii UCP1619 was
obtained from the culture bank of the Centre for Research in Environ-
mental Sciences of the Catholic University of Pernambuco, Brazil (UNI-
CAP) and used to produce the BC films.
To maintain the viability of the microorganism and its cultivation,
a standard HS medium adapted by Hungund and Gupta (2010) and
Gomes et al. (2013) was used. The HS medium was composed of
20.00 g.L‐1 of glucose, 5.00 g.L‐1 of peptone, 5.00 g.L‐1 of yeast extract,
2.70 g.L‐1 of disodium phosphate, and 1.15 g.L‐1 of citric acid (pH 5).
For preservation of the microorganism, 15.00 g.L‐1 of agar was added
to the medium. All media were autoclaved at 121 ± 1 °C for 15 min.
Pre-inoculum and inoculum
The pre‐inoculum consisted of growing the microorganism at
30 ± 2 °C for 48 h under static conditions in 100 mL of HS medium
in 250‐mL Erlenmeyer flasks. For inoculation, 3% (v/v) of the pre‐
inoculum was added to 200 mL of HS liquid medium in 500‐mL Erlen-
meyer flasks under the same conditions.
Acquisition of extracts
Concord grape (Vitis labrusca) and red cabbage (Brassica oleracea
var. capitate f. rubra) extracts were obtained with an infusion method.
Concord grape skin was weighed and added to a 1‐L beaker with pre-
viously heated distilled water (90 °C) for 10 min at a concentration of
375.00 g.L‐1. The resulting solution had a pH of 4. After filtration, the
extract was stored at 4 °C in an amber bottle until further use. For the
red cabbage extract, the leaves were initially washed with distilled
water and cut before being subjected to the infusion method. The
experimental conditions were the same as those described for the
grape extract. This process resulted in an extract with a concentration
of 375.00 g.L‐1 and pH 6.
BC production in alternative culture media
The production of BC membranes occurred in standard HS and
modified HS media with different proportions of concord grape skin
and red cabbage extracts. The bacterial cell suspension [3% (v/v)]
was transferred to 100 mL of the alternative production media in a
500‐mL Schott flask under static conditions at 30 ± 2 °C for 7 days.
Table 1 shows the composition and pH of each production medium
used to obtain the BC. All experiments were performed in triplicate.
BC membrane washing and purification
BC membranes were collected, washed in distilled water, and puri-
fied to eliminate bacterial cells trapped in the membranes. The purifi-
cation process was performed as described by Amorim et al. (2020)
and Suryanto et al. (2019): the membranes were immersed in NaOH
solution (0.1 mol.L‐1) at 90 °C for 20 min, then washed with deionised
water until reaching pH 7.
Sample sterilisation by gamma irradiation
After production, and without any treatment, the BC membranes
were subjected to gamma sterilisation with irradiation at a dose of
25 kGy (Paula et al., 2019) for 15 h and 35 min. Irradiation was per-
formed in the presence of air at a temperature of 25 ± 3 °C with the
aid of a 60Co sealed solid source radiator (rate: 1,604 kGy.h−1) with
Gammacell 220 Excel MDS Nordion equipment at the Laboratory of
Metrology of Ionising Radiation of the Department of Nuclear Energy
of the Federal University of Pernambuco (LMRI‐DEN/UFPE).
H. Almeida do Nascimento et al. Current Research in Biotechnology 4 (2022) 119–128

Table 1
Composition of modified media used in the production of bacterial cellulose.

Composition Extract (% v/v) Hestrin-Schramm medium (% v/v) pH Label

Standard medium 0 100 5.0 HS

Concord grape skin 100 0 4.0 G100
50 50 4.4 G50
25 75 4.9 G25
Purple cabbage 100 0 6.0 C100
50 50 5.3 C50
25 75 5.8 C25

Water holding capacity and porosity Scanning electron microscopy

Wet BC membranes were weighed and dried in a benchtop oven at The membranes were dried and covered with a conductive gold
50 ± 1 °C for complete water removal (until a constant weight was film. SEM was performed with a VEGA3 Tescan electron microscope
reached). The water holding capacity (WHC) percentage was deter- operating at10 Kv.
mined with Eq. (1).
Results and discussion
ðMean mass of wet membranesðgÞ  Mean mass of dry membranes ðgÞÞ
WHCð%Þ ¼
ðMean mass of wet membranesðgÞÞ
BC membrane production
ð1Þ

The surface porosity of the BC membranes was determined with Eq.
(2) (Ding et al., 2016).
ðW w  W d Þ
Porosity ¼ ð Þx100%
dxDxA
in which Ww and Wd are the wet and dry membrane weights (g),
respectively, d is water density (1 g.cm−3), D is membrane thickness
(cm), and A is membrane area (cm2).
Fourier transform infrared spectroscopy
The samples were dehydrated and kept in a desiccator with silica
gel at room temperature (25 ± 3 °C), then digitised in a Bucker FTIR
spectrometer (Equinox 55 Model, Bruker Co., Ettlingen, Germany)
with a horizontal attenuated total reflectance device through a plate
of crystal cells (45° ZnSe; 80 × 10 mm thickness: 4 mm) (PIKE Tech-
nology Inc., Madison, WI, USA). The scanning range was 4000 to
400 cm−1, with 32 scans and a resolution of 4 cm−1.
The different supplemented culture media were evaluated on the
basis of BC yield (wet and dry weight), as shown in Table 2. Character-
istic colour changes were found for each BC membrane, corresponding
to the colours of the extracts used in the production media (Fig. 1). The
ð2Þ thickness of the membranes varied in accordance with the yields
(Table 2), because all samples had the same diameter (Fig. 2).
Only one pure extract medium was able to obtain a membrane
(G100), resulting in minimum production. In contrast, C100 had no
cellulose production. These results confirmed the dependence of BC
production on the availability and type of carbon source in the culture
medium, as reported by Costa et al. (2017). The absence of BC produc-
tion in the C100 medium might also have been explained by the lack of
nutrients (particularly the availability of carbon sources).
The best alternative medium was C50, with an average yield of
155.55 ± 3.90 g.L‐1 for hydrated cellulose and 3.76 ± 0.38 g.L‐1 for
dried cellulose, with a production time of 7 days. The other results
were compatible to those found in previous studies with alternative
media involving plant extracts for BC production (Amorim et al.,
2019; Costa et al., 2017). In general, the results revealed a consider-
able decrease in the cost of the production process.

X-ray diffraction Water holding capacity and porosity

XRD patterns were measured with a Rigaku diffractometer with a The WHC and porosity values of the BC membranes were deter-
copper tube, voltage of 40 kVe, and a current of 20 mA on the 2θ scale, mined after 7 days of fermentation. The results are shown in Table 3.
with a range of 10° to 30° and scanning speed of 0.5°.min−1. Eq. (3) The BC membranes had a high WHC, with values around 98%. Such
was used to calculate the crystallinity index (CI) on the basis of the dif- behaviour has been described in the literature, thus demonstrating
ference between the highest and lowest intensity peaks, which corre- Table 2
sponded to the crystalline (Ic) and amorphous (Ia) phases, Yields expressed in wet and dry weight according to culture media.
respectively (Segal et al., 1959).
Culture media Wet bacterial cellulose Dry bacterial cellulose
weight weight
Ic  Ia (g.L-1) (g.L-1)
CI ¼ x100% ð3Þ
Ic μ±σ μ±σ

Hestrin-Schramm 120.55 ± 2.41 1.90 ± 0.27

(HS)
Grape 100% (G100) 84.45 ± 2.35 0.92 ± 0.13
Thermogravimetric analysis
Grape 50% (G50) 122.89 ± 3.74 1.81 ± 0.32
Grape 25% (G25) 134.45 ± 1.92 2.68 ± 0.51
Thermal stability was determined with TGA. Approximately 8 mg Cabbage 100% – –
of each sample was weighed, then scanned from 30 °C to 600 °C under (C100)
a nitrogen atmosphere with a flow rate of 20 mL.min−1 to avoid Cabbage 50% (C50) 155.55 ± 3.90 3.76 ± 0.38
Cabbage 25% (C25) 149.15 ± 3.81 2.94 ± 0.40
thermo‐oxidative degradation of the samples (heating rate: 10 °C.
min−1). A Mettler Toledo TGA 2 Star System analyser was used. μ = mean; σ = standard deviation.

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H. Almeida do Nascimento et al.
Fig. 1. Aspects of wet bacterial cellulose (BC) membranes produced with different culture media.
that the water content of BC membranes is high regardless of the pro-
duction time and medium (Amorim et al., 2019; Galdino et al., 2020;
Costa et al., 2017). Furthermore, the porosity values were compatible
with the obtained yields. A decrease in porosity occurred with increas-
ing membrane thickness, that is, when a higher yield value was
obtained. This behaviour was expected, because greater thickness is
associated with the increase in the concentration of the fibres that
form the membranes.
X-ray diffraction
XRD was used to evaluate the crystalline structure and CI of the BC
membranes produced with different media before and after exposure
to gamma irradiation (G). The CI values are shown in Table 4.
The BC membrane produced with standard HS medium achieved
the highest CI and maintained its value after undergoing irradiation,
with a difference of 0.33%. A slight CI decrease was observed in both
samples with 25% extract content (G25 and C25: 5.19% and 0.45%,
respectively). Similar results have been described in the literature
(Hwang et al., 2021). In contrast, sample C50 showed a 39.18%
decrease in the CI. Further investigations are needed to explain the
increase in the amorphous region.
XRD profiles were generated to determine the composition of each
membrane (Fig. 3). In all samples, two peaks with 2θ angles around
14.5° and 23° were observed. These corresponded to crystalline planes
(1 1 0) and (2 0 0), respectively, thus demonstrating that all obtained
membranes, regardless of the culture medium, were type I cellulose
(Abdelraof et al., 2019). A peak intensity decrease was also observed
after gamma irradiation in the samples obtained with the extracts, thus
confirming the decrease in CI presented in Table 4. The diffractograms
of the irradiated samples were similar to those before irradiation, thus
indicating that treatment did not create new crystalline planes of sym-
metry, as previously reported by Paula et al. (2019).
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Current Research in Biotechnology 4 (2022) 119–128
Fourier transform infrared spectroscopy
Fig. 4 shows similar FTIR spectra for the membranes produced with
different culture media before and after the irradiation process. The
FTIR spectra showed the main characteristic bands of BC. The band
at 3336 cm−1 corresponded to the vibrational stretching of the hydro-
xyl groups (OH). The band at 2895 cm−1 was attributed to the asym-
metrical vibrational stretching of methylene bridges (–CH2‐). The
band at 2839 cm−1 corresponded to the symmetrical vibrational
stretching of the methyl groups (–CH3). Bands in the regions of
1427 cm−1 and 1315 cm−1 corresponded to C‐OH and CH bonds.
The bands in the regions of 1160 cm−1 and 1109 cm−1 corresponded
to the stretching of the CO and C‐OH groups. The band at 1055 cm−1
was attributable to the elongation of the COC and C‐OH bonds in car-
bohydrates (Pourjavaher et al., 2017; Revin et al., 2018; Khan et al.,
2020).
Fig. 4a and 4b also show a band in the region of 1649 cm−1 that
corresponds to the vibration of the H‐O‐H bond, according to the water
content of the membranes (Khan et al., 2020). In the irradiated sam-
ples (Fig. 4c and Fig. 4d), this band shifted slightly to 1642 cm−1,
but this behaviour did not cause any structural changes. According
to the results of a study by Pourjavaher et al. (2017), the aforemen-
tioned bands are also attributable to the stretching vibration of the aro-
matic ring (C = C) of phenolic compounds, such as anthocyanin, a
flavonoid responsible for the red, purple, and blue colours of various
types of plants and fruits, such as grapes and red cabbage.
Regarding the sample produced with pure grape extract (G100),
the band in the region of 1716 cm−1 corresponded to the stretching
of the carbonyl group (C = O), thus indicating the strong presence
of polysaccharides, commonly found in the cell walls of plant tissues,
particularly fruits, in addition to the presence of some phenolic acids
(Lu and Hsieh, 2012; Bueno et al., 2017).
H. Almeida do Nascimento et al. Current Research in Biotechnology 4 (2022) 119–128

Fig. 2. Culture media used for bacterial cellulose (BC) production.

Table 3 Table 4
Water holding capacity (WHC) and porosity of bacterial cellulose membranes Crystallinity indices of bacterial cellulose membranes before and after exposure
obtained with different production media. to gamma irradiation, obtained with different production media.

Water holding capacity (%) Porosity (%) μ ± σ Non-irradiated Crystallinity Irradiated membranes Crystallinity
μ±σ membranes index (%) μ ± σ index (%) μ ± σ
Culture media
Hestrin- 75.61 ± 0.37 Hestrin-Schramm – 75.36 ± 0.27
Hestrin-Schramm (HS) 98.42 ± 0.34 76.55 ± 0.51 Schramm Gamma (HS – G)
Grape 100% (G100) 98.94 ± 0.39 89.84 ± 0.53 (HS)
Grape 50% (G50) 98.53 ± 0.31 86.80 ± 0.50 Grape 100% 72.95 ± 0.83 Grape 100% – Gamma 56.37 ± 0.48
Grape 25% (G25) 98.01 ± 0.17 85.02 ± 0.39 (G100) (G100 – G)
Cabbage 50% (C50) 97.58 ± 0.23 61.20 ± 0.45 Grape 50% 60.02 ± 1.50 Grape 50% – Gamma 47.35 ± 1.52
Cabbage 25% (C25) 98.03 ± 0.18 77.61 ± 0.39 (G50) (G50 – G)
Grape 25% 59.08 ± 0.91 Grape 25% – Gamma 56.01 ± 0.21
μ = mean; σ = standard deviation. (G25) (G25 – G)
Cabbage 50% 68.70 ± 0.72 Cabbage 50% – 41.78 ± 1.99
(C50) Gamma (C50 – G)
The results demonstrated that sterilisation with gamma irradiation Cabbage 25% 44.22 ± 0.65 Cabbage 25% – 44.02 ± 0.84
(C25) Gamma (C25 – G)
at 25 KGy caused no significant structural changes in the polymer
chains. Similar data have been reported by Salari et al. (2021), μ = mean; σ = standard deviation.
Hodder et al. (2019), and Li et al. (2011).

Thermogravimetric analysis
Dried BC samples were used to determine thermal stability. The
curves before and after exposure to gamma irradiation are shown in
Fig. 5. All samples showed two stages of mass loss before sterilisation
by gamma irradiation. The first stage refers to the evaporation of the
remaining water absorbed in the BC membrane, which occurred at
90 °C with the HS medium and in the temperature range of
112–180 °C for the samples obtained with grape and cabbage extracts.
This increase might have been due to the degradation of the con-
stituents of the extracts.

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Fig. 3. X-ray diffractograms of bacterial cellulose (BC) membranes produced with different media before (HS, G100, G50, G25, C50, and C25) and after exposure
to gamma irradiation (HS-G, G100-G, G50-G, G25-G, C50-G, and C25-G).

The second loss of mass was associated with the thermal degrada- C, which might have been associated with higher concentrations of
tion of the BC, which occurs in a range of higher temperatures polysaccharides and phenolic compounds in the grape extract.
(260–365 °C). In this second phase, degradation, depolymerisation, According to Salari et al. (2021), the effects of gamma irradiation
dehydration, and decomposition of the glucose monosaccharide units on the thermal properties of biopolymers are directly associated with
occurred, and carbon residues consequently formed. These results the type of biopolymer and the dosage of radiation used in the process.
are similar to those described in the literature (Costa et al., 2019; Thus, a change can be observed in the maximum temperature
Gao et al., 2019). One criterion for evaluating the material degradation achieved.
process is the maximum temperature reached. For irradiated BC mem-
branes, a small increase in the maximum degradation temperature,
with respect to that of non‐irradiated samples, was observed, with val-
Scanning electron microscopy
ues around 250–360 °C.
An exception occurred for G100‐G, in which significant changes
The SEM images of the dry BC membranes before and after sterili-
were found with the emergence of different stages. Such stages can
sation with gamma irradiation had similar aspects, as shown in Fig. 6.
be explained by structural changes to the BC after gamma irradiation.
Characteristic morphological aspects of BC were observed, such as an
This sample had a lower fibre concentration in its structure, thus sug-
ultra‐fine nanofibrillary reticulated structure with irregular fibre dis-
gesting that the dose of 25 kGy was too high for this thickness, thereby
tribution arranged in a three‐dimensional porous network, as also
causing changes in the polymer. The second and third stages (temper-
reported in other studies (Wang et al., 2021; Galdino et al., 2020).
ature ranges around 225 °C and 263 °C, respectively) may be associ-
In some micrographs, several bacterial cells in the shape of bacilli were
ated with the degradation of the constituents of the extract. Two
seen enveloped by the nanofibrillar structure. This finding was
additional stages occurred at temperatures around 349 °C and 580 °
expected, because fibres are excreted from these cells.

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H. Almeida do Nascimento et al. Current Research in Biotechnology 4 (2022) 119–128

Fig. 4. Fourier transform infrared spectra of bacterial cellulose (BC) membranes produced with different media before and after exposure to gamma irradiation.

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H. Almeida do Nascimento et al. Current Research in Biotechnology 4 (2022) 119–128

Fig. 5. Thermogravimetric curves of bacterial cellulose (BC) membranes produced with different media before and after exposure to gamma irradiation.

Fig. 6. Scanning electron microscopy images with 10.0 kx of bacterial cellulose (BC) membranes produced with different media before and after exposure to
gamma irradiation.

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H. Almeida do Nascimento et al.
Conclusion
This work involved experimental evaluation of bacterial cellulose
production by using standard HS medium and alternative media for-
mulated with different compositions of concord grape and red cabbage
extracts. The effects of gamma irradiation on the polymer were also
assessed. The medium with the best production yield was C50, which
enabled a considerable decrease in production costs. Characteristics
associated with semi‐crystalline, chemical, and morphological struc-
tures, as well as the thermal stability of the obtained BC membranes,
were evaluated. The XRD analysis indicated that irradiation did not
create new planes of symmetry. However, decreases were found in
the CI values. The FTIR and SEM data demonstrated no changes in
the BC polymeric structure. Regarding thermal stability, the BC sam-
ples exhibited different behaviours with each production medium.
After irradiation, a small decrease occurred in the maximum degrada-
tion temperatures of most samples. The findings suggest that sterilisa-
tion with gamma irradiation at a dose of 25 kGy is a favourable
technique, because it did not cause significant changes in the proper-
ties of the BC membranes studied.
CRediT authorship contribution statement
Helenise Almeida do Nascimento: Data curation, Formal analy-
sis, Investigation, Writing – review & editing. Julia Didier Pedrosa
Amorim: Conceptualization, Data curation, Formal analysis, Investiga-
tion, Methodology, Visualization, Writing – review & editing, Formal
analysis. Cláudio José Galdino da Silva Júnior: Writing – review
& editing, Methodology, Formal analysis. Alexandre D'Lamare Maia
de Medeiros: Writing – review & editing, Formal analysis. Andréa
Fernanda de Santana Costa: Conceptualization, Supervision.
Daniella Carla Napoleão: Conceptualization, Writing – review & edit-
ing. Glória Maria Vinhas: Conceptualization, Methodology. Leonie
Asfora Sarubbo: Visualization, Writing – review & editing, Funding
acquisition, Resources, Supervision.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgements
The authors gratefully acknowledge IATI, UNICAP, and UFPE for pro-
viding all necessary facilities to perform the research.
Funding
This work was supported by the Brazilian fostering agencies Coor-
denação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES
[Coordination for the Advancement of Higher Education Personnel]
– Finance Code 001), Conselho Nacional de Desenvolvimento Cientí-
fico e Tecnológico (CNPq [National Council for Scientific and Techno-
logical Development]), Universidade Federal de Pernambuco (UFPE),
Escola Icam Tech–Universidade Católica de Pernambuco (UNICAP)
and Instituto Avançado de Tecnologia e Inovação (IATI)
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