Rein o cemen o soy p o ein-based bioplas ics as po en ial sus ainable
packaging solu ions
Ca men Ma ía G anados-Ca e a
a,*
, Daniel Cas o-C iado
a
, Me cedes Jim´
enez-Rosado
b
,
Albe o Rome o
a,*
, Víc o Manuel Pe ez-Puyana
c
a
Depa men o Chemical Enginee ing, Facul y o Chemis y, Uni e si y o Se ille 41012, Se ille, Spain
b
Depa men o Applied Chemis y and Physics, Facul y o Biological and Ambien al Sciences, Uni e si y o Le´
on, 24071, Le´
on, Spain
c
Depa men o Enginee ing and Ma e ials Science and T anspo a ion, Uni e si y o Se ille 41092, Se ille, , Spain
ARTICLE INFO
Keywo ds:
Bioplas ics
Ag i- ood was e
Soy p o ein
C osslinking
Injec ion moulding
ABSTRACT
Due o he subs an ial amoun o plas ic was e in he en i onmen , scien is s a e seeking new al e na i es o
adi ional plas ics. Bioplas ics a e conside ed o be impo an in add essing his issue despi e hei signi ican
d awbacks, such as poo mechanical p ope ies and highe cos s. In o de o educe hei p ice, ag i- ood was e
and by-p oduc s can be used as aw ma e ials (e.g., soy p o ein), p omo ing a ci cula economy; and by inco -
po a ing di e en ein o cemen me hods, i is possible o de elop ma e ials wi h imp o ed mechanical and
ba ie p ope ies. The aim o his wo k is o imp o e he p ope ies o soy p o ein/glyce ol injec ed bioplas ics
by inco po a ing di e en biopolyme s (gela in and saccha ose) o applying di e en c osslinking me hods
(physical, chemical o enzyma ic c osslinking h ough he mal ea men , glyoxal o ansglu aminase, espec-
i ely). These ma e ials we e e alua ed by physicochemical, mechanical, and unc ional es s. The esul s
con i med an imp o emen in he mechanical p ope ies o he ein o ced p o ein-based bioplas ics, showing an
inc ease in hei s i ness and a dec ease in hei de o mabili y, educing hei capaci y o abso b wa e . In any
case, hese esul s suppo he modi ica ion o he p ope ies compa ed o he e e ence sys ems.
1. In oduc ion
Ag icul u al and ood was e, commonly known as ag i- ood was e,
has a nega i e impac on ou daily li es as well as on he en i onmen ,
socie y and economy. Globally, a ound 1.3 billion onnes o ood a e los
be o e human consump ion, which ep esen s one- hi d o he o al
p oduc ion, leading o social impac s such as nu ien loss and wo ld
hunge (Capanoglu e al., 2022; Ma ei e al., 2021). Howe e , he al-
o isa ion o ag icul u al and ood was es and by-p oduc s can be a po-
en ial solu ion o achie e a g een ci cula economy (J˜
ogi and Bha ,
2020; Paini e al., 2022; Valencia e al., 2021). Hence, hese was es can
be ans o med in o alue-added p oduc s, such as bioplas ics, o
add ess he la ge numbe o plas ics accumula ed in ecosys ems
(Ben-O hman e al., 2020; Tsang e al., 2019; Valencia e al., 2021).
Plas ics a e used in a wide ange o daily applica ions. The packaging
indus y is pa icula ly no ewo hy, accoun ing o 39.7 % o o al de-
mand, making his sec o one o he la ges consume s due o he in-
c ease in pu chases o manu ac u ed and packaged p oduc s (Fog
Jacobsen e al., 2022; Foschi and Bonoli, 2019). Speci ically, in he case
o ood packaging, which is exposed o ex e nal de e io a ion me hods
such as mechanical o ces o wa e apou , among o he s, one o he
main conside a ions is he lowe pe meabili y o a oid con ac wi h
oxygen and wo k as a ba ie (Ha nka nsuja i e al., 2021; Me ino e al.,
2022; San ana e al., 2021). Nowadays, in pa icula , app oxima ely 95
% o he plas ics used a e de i ed om non- enewable esou ces (Shaikh
e al., 2021), a e single-used and encou age he accumula ion o was e in
land ills, con ibu ing o almos 50 % o he o al global plas ic was e
(Raha diyan e al., 2023; Shang e al., 2023). The main d awback o
hese ma e ials is ha hey accumula e in he ecosys ems and pe sis o
cen u ies wi hou deg ada ion due o hei high esis ance o mic obial
deg ada ion (La agnolo e al., 2024; Lim e al., 2023; Yin and Woo,
2024). The e o e, scien is s a e ocusing on he de elopmen o new
biodeg adable and eco- iendly plas ics ha can eplace con en ional
plas ics (Kuma i e al., 2023; Pascoe O iz, 2023), ocusing on he
applica ion o hese biodeg adable plas ics in ood packaging and ag i-
cul u al sec o s (Shaikh e al., 2021). Bioplas ics ha e been de eloped
* Co esponding au ho s.
E-mail add esses: [email p o ec ed] (C.M. G anados-Ca e a), [email p o ec ed] (D. Cas o-C iado), [email p o ec ed] (M. Jim´
enez-Rosado), al ome o@
us.es (A. Rome o), [email p o ec ed] (V.M. Pe ez-Puyana).
Con en s lis s a ailable a ScienceDi ec
Fu u e Foods
jou nal homepage: www.else ie .com/loca e/ u o
h ps://doi.o g/10.1016/j. u o.2024.100524
Recei ed 13 Oc obe 2024; Recei ed in e ised o m 13 No embe 2024; Accep ed 11 Decembe 2024
Fu u e Foods 11 (2025) 100524
A ailable online 14 Decembe 2024
2666-8335/© 2024 The Au ho s. Published by Else ie B.V. This is an open access a icle unde he CC BY-NC-ND license (
h p://c ea i ecommons.o g/licenses/by-
nc-nd/4.0/ ).
using di e en aw ma e ials such as s a ch, suga cane o co n (Ali
e al., 2023; Jayakuma e al., 2023; Kong e al., 2023), and me hods
such as cas ing, he momoulding, 3D-p in ing, among o he s (Ado na
e al., 2022; Ca ajal-Pi˜
ne o e al., 2019; Dey e al., 2023). Cu en ly,
hey a e conside ed as an impo an way o achie e sus ainable de el-
opmen goals, such as educing eliance on ossil uels, minimising he
use o oxic chemicals, and mo ing owa ds ecycling (Ahsan e al.,
2023).
Howe e , one majo d awback o bioplas ics is hei cos , which
anges om 1.14 o 21.50
€
pe kilog am, compa ed o 0.57 o 1.59
€
pe
kilog am o con en ional plas ics. This cos di e ence makes bio-
plas ics less compe i i e and limi s hei applicabili y (Gong e al., 2024;
Jim´
enez-Rosado e al., 2020; Ka an e al., 2019; Pa ia e al., 2024). In
o de o educe hei cos , ag i- ood was e can be used as aw ma e ial
wi hou comp omising ood sa e y (Gong e al., 2024; Mo one e al.,
2019). Ag i- ood was e and by-p oduc s a e ich in p o ein (Bagnani
e al., 2024; J˜
ogi and Bha , 2020), which p omo es i s alo isa ion o
de elop bioplas ics as a high-added alue p oduc (Cas o-C iado e al.,
2024). In his con ex , soy p o ein isola e (SPI) is an encou aging aw
ma e ial due o i s a ou able p ope ies such as biodeg adabili y o
good abso p ion capaci y (Cuad i e al., 2017; Rani e al., 2021; Song
e al., 2011; Yan e al., 2021), as well as i s hyd ophilic cha ac e (as a
consequence o he high p esence o acidic amino acids o aspa ic acid
and glu amic acid in i s composi ion) (Jim´
enez-Rosado e al., 2019;
Lamaming e al., 2021; Yamada e al., 2020).
On he o he hand, ano he d awback associa ed wi h bioplas ics is
hei poo mechanical p ope ies and poo wa e esis ance (Lusiana
e al., 2019; O hman e al., 2021; Zhang e al., 2022), being necessa y o
i s applica ion in ood packaging an imp o emen o he p ope ies since
he momen he ood wi hin i ill he end o he consump ion (Me ino
e al., 2022). Howe e , hese d awbacks can be o e come by using
di e en ein o cemen s a egies (Awadhiya e al., 2016; Ishak e al.,
2020). Chemical c osslinking can be used o impa mechanical s i ness
o p o ein s uc u es, and his can be de eloped by nume ous agen s
such as aldehydes (e.g. glyoxal, glu a aldehyde o o maldehyde) o
acids (e.g. ci ic acid) due o he exis ence o mul iple mul i unc ional
g oups (´
Al a ez-Cas illo e al., 2021; Ma qui´
e, 2001), highligh ing
glyoxal due o i s abili y o p oduce bioplas ics wi h excellen mechan-
ical and he mal p ope ies (Ishak e al., 2020; Z´
a a e-Ramí ez e al.,
2014). On he o he hand, physical c osslinking, such as hea ea men ,
can be used o p omo e he o ma ion o co alen bonds and inc ease
ensile modulus and s eng h by educing elonga ion (Du e al., 2016;
Jim´
enez-Rosado e al., 2020; Pe ez-Puyana e al., 2022; Zhang e al.,
2023); mo eo e , his me hod highligh s o i s acili y and he modi i-
ca ion ha p omo es in wa e abso p ion capabili y and he iscoelas ic
p ope ies o he ma ix (´
Al a ez-Cas illo e al., 2018; Xie e al., 2022).
Ano he al e na i e can be he inco po a ion o addi i es, which can
also imp o e ce ain unc ional p ope ies (Kong e al., 2023). Fo
example, gela in is a p o ein ha can inc ease mechanical esis ance,
ensile s eng h, hyd ophilici y, swelling capaci y and anspa ency, as
shown in p e ious s udies (Galus, 2017). Simila ly, o he s udies ha e
ocused on he inco po a ion o o he biodeg adable polyme s, such as
p opylene glycol algina e (PGA), which p omo es he op imisa ion o
wa e esis ance h ough he o ma ion o co alen complexes (Zhang
and Se en i, 2020). Polysaccha ides such as chi osan (CH) can also be
added due o hei hyd ophobici y, high in e acial ension and hei
imp o emen o he wa e ba ie beha iou (Haghighi e al., 2019).
Mo eo e , disaccha ides, such as saccha ose, can p omo e an inc ease in
in e molecula in e ac ion and ensile s eng h, as seen in p e ious
s udies (Wang e al., 2022). Finally, he inco po a ion o enzyma ic
c osslinke s, such as ansglu aminase, p omo es a highe ensile
s eng h and su ace hyd ophobici y (Mohammad Zadeh e al., 2018;
Wang, 2022).
The e o e, his s udy ocused on he e alua ion o di e en me hods
o ein o cing soy p o ein-based bioplas ics by inco po a ing di e en
biopolyme s (gela in and saccha ose) o applying di e en c osslinking
me hods (physical, chemical o enzyma ic c osslinking h ough he mal
ea men , glyoxal o ansglu aminase, espec i ely). La e , he physi-
cochemical, he momechanical and unc ional p ope ies we e s udied.
Finally, he main no el y o his s udy is he compa ison be ween he
esul s ob ained by di e en c osslinking me hods on soy-based
bioplas ics.
2. Ma e ials and me hods
2.1. Ma e ials
Soy p o ein isola e (SPI, 91 w % p o ein and 6 w % mois u e) used as
aw ma e ial was supplied by P o ein Technologies In e na ional
(SUPRO 500E, Belgium). Food gela in (G, 90 w % p o ein) and sac-
cha ose (S, pu i y ≥95 %), used as ein o cing biopolyme s, we e sup-
plied by Manuel Riesgo L d. (Spain) and Sigma Ald ich S.A. (Ge many),
espec i ely. Glyce ol (Gly), which was p o ided by Pan eac Química
L d. (Spain), was used as plas icise . Finally, glyoxal (Glyx) and ans-
glu aminase (Tgase, enzyma ic ac i i y o 100 uni s/g) we e p o ided by
Pan eac Química L d. (Spain) and BDF Ing edien s (P obindTX-Tgasa,
Spain), espec i ely.
2.2. P epa a ion o bioplas ics
The bioplas ics we e p ocessed by injec ion moulding, which consis s
o wo s ages: a mixing s age and an injec ion moulding s age. Fi s , a
wo-blade coun e - o a ing heome e mixe Polylab QC (The moHaake,
Ge many) was used o homogenise he di e en aw ma e ials p esen in
he bioplas ics. In his case, he condi ions imposed we e an angula
speed o 50 pm o 10 min unde adiaba ic condi ions s a ing om
oom empe a u e (25 ±2 ◦C). Du ing mixing, he o que and empe -
a u e gene a ed we e ollowed and di e en p o ein/plas icise /addi-
i es a ios we e analysed o selec a sui able p opo ion, as shown in
Table 1.
Subsequen ly, an injec ion moulding s age was ca ied ou using a
MiniJe Pis on Injec ion Moulding Sys em (The moHaake, Ge many), in
which he p e iously ob ained blends we e placed in a cylind ical p e-
chambe a 40 ◦C and hen o ced h ough a nozzle using a plunge
(600 ba o 20 s) o low in o he ca i ies o a mould ( ec angula , 60 ×
10 ×1 mm
3
) a 90 ◦C, whe e he bioplas ics we e subjec ed o a
densi ica ion s age a 200 ba o 300 s (Jim´
enez Rosado, 2022). I is
wo h no ing ha he sys ems con aining saccha ose we e p ocessed a
70 ◦C wi h injec ion and pos -injec ion p essu es o 500 and 300 ba ,
espec i ely, o a oid saccha ose deg ada ion and enhance wa e ab-
so p ion o he sys ems (Abd-El ahman and Ahmed, 2009;
Jim´
enez-Rosado e al., 2021).
Fu he mo e, he bioplas ics subjec ed o physical c osslinking un-
de wen an addi ional s ep in a con en ional o en (Memme , Ge many)
Table 1
Composi ion o he di e en bioplas ic ma ices.
Sys em Pe cen age (w %) Rein o cemen
SPI Gly Tgase Glyx S G
Re 50 50 – – – – –
G-5 45 50 – – – 5 Adi i e
G-10 40 50 – – – 10 Adi i e
G-20 30 50 – – – 20 Adi i e
S-5 47.5 47.5 – – 5–Adi i e
S-10 45 45 – – 10 –Adi i e
S-20 40 40 – – 20 –Adi i e
HT-50 50 50 – – – – Hea ea men
HT-120 50 50 – – – – Hea ea men
Glyx-1 49.5 49.5 –1– – Chemical
Glyx-3 48.5 48.5 –3– – Chemical
Tgase-0.1 49.95 49.95 0.1 – – – Enzima ic
Tgase-0.2 49.90 49.90 0.2 – – – Enzima ic
C.M. G anados-Ca e a e al.
Fu u e Foods 11 (2025) 100524
2
o 24 h a di e en empe a u es (50 and 120 ◦C) (´
Al a ez-Cas illo
e al., 2018; Fe n´
andez-Espada e al., 2016).
2.3. Cha ac e iza ion o samples
2.3.1. Physicochemical p ope ies
2.3.1.1. Fou ie ans o m in a ed spec oscopy (FTIR). A Fou ie
T ans o m In a ed Spec oscopy (FTIR) analysis was conduc ed using a
Hype ion 100 spec opho ome e by B uke , USA. This de ice was u i-
lized o iden i y he a ious chemical bonds p esen in he bioplas ic
ma ices. The spec a we e depic ed in a wa enumbe spec um anging
om 4000 o 400 cm
−1
and hen p ocessed using Jasco Spec a Man-
age TM so wa e, e sion 2.
2.3.1.2. Wa e con ac angle (WCA). The wa e con ac angle was
measu ed using an op ical ensiome e (A en ion TL 10, KSV, Finland)
and he sessile d op me hod. A 2 µL d op o milli-Q g ade wa e was
placed on he bioplas ic samples’ su ace, and he d ople ’s shape was
analysed o 10 s. The wa e con ac angle was measu ed on bo h sides o
he d ople , and he a e age alue was calcula ed.
2.3.2. Mechanical p ope ies
2.3.2.1. Dynamic lexu e es . Dynamic-mechanical analyses we e ca -
ied ou o e alua e he linea iscoelas ic esponse o he di e en
bioplas ics. Two es s we e pe o med in lexu al mode using a
mechanical-dynamic analyse RSA3 (TA Ins umen s, USA) wi h a dual
can ile e geome y a oom empe a u e (25 ±2 ◦C):
•S ain sweep es s we e pe o med be ween 0.002 and 1 % o s ain
a a cons an equency o 1 Hz o de e mine he linea iscoelas ic
ange (LVR), whe e he elas ic and iscous moduli emain indepen-
den o he applied de o ma ion. F om hese es s, he c i ical
de o ma ion (%) was de e mined, which ma ks he limi o he LVR.
•F equency sweep es s we e de eloped be ween 0.02 and 20 Hz,
applying a cons an s ain wi hin he iscoelas ic ange. F om his
es , he elas ic (E’) and iscous (E’’) moduli and he loss angen ( an
(δ)=E’’/E’) we e ob ained in he es ablished equency ange. F om
hese es s, E’
1
and an (δ)
1
we e used as he co esponding alues o
E’ and an (δ) a 1 Hz o acili a e compa ison be ween di e en
sys ems.
2.3.2.2. Tensile es s. Tensile es s un il b eakage we e ca ied ou in an
Insigh 10 kN Uni e sal Tes ing Machine (MTS, USA) o s udy he
s eng h o he bioplas ics. The wo king condi ions o he ensile es
we e se acco ding o he ISO 527 2.2 (2012) s anda d. In his es , he
bioplas ics we e subjec ed o an inc easing axial o ce a a a e o 5 mm/
min. The applied s ess is measu ed by he s ain o he bioplas ic un il i
b eaks. This es p o ides da a on Young’s modulus, maximum s ess
and s ain a b eak.
2.3.3. Func ional p ope ies
2.3.3.1. Wa e up ake capaci y and soluble ma e loss. Wa e up ake
capaci y (WUC) and soluble ma e loss (SML) we e ca ied ou ac-
co ding o he p ocedu e p e iously used by Jim´
enez-Rosado e al.
(Jim´
enez-Rosado e al., 2019). Thus, he di e en bioplas ics we e
imme sed in a closed essel wi h 30 mL o dis illed wa e o 24 h Then,
WUC was calcula ed using Eq. (1), whe e w
2
e e s o he weigh o he
bioplas ic a e wa e abso p ion and w
3
e e s o he weigh o he d y
bioplas ic a e he abso p ion (d ied in an o en a 120 ◦C o 2 h).
WUC (%) = w2−w3
w3
⋅100 (1)
Finally, SML can be calcula ed using Eq. (2), in which w
1
e e s o he
weigh o he d y bioplas ic be o e he es .
SML (%) = w1−w3
w1
⋅100 (2)
2.3.3.2. Biodeg adabili y. The biodeg adabili y o he sys ems which
inco po a e gela in as a blending componen , was measu ed by bu ying
he bioplas ics a oom empe a u e in a compos ing medium (2:1
c opland: compos , he same a io o ine /o ganic ma e ials as speci ied
by ISO 20,200 (Paulson e al., 2001)). Speci ically, hese sys ems we e
chosen due o he he e ogenei ies p esen in hei bioplas ic samples o
e alua e i he e was any ela ion wi h biodeg ada ion.
A leas h ee ec angula bioplas ics om each sys em we e e alu-
a ed by digging hem up o isual e alua ion on di e en days, and
pho og aphs we e aken o he sys ems o his pu pose. The es was
conside ed ended when no po ion la ge han 1 mm o he bioplas ic
could be unea hed (es ablishing he o al biodeg ada ion ime).
2.4. S a is ical analysis
The di e en measu emen s we e pe o med a leas in iplica e. In
o de o ob ain ep esen a i eness in he sys ems, a s a is ical analysis
was ca ied ou using a compa ison o means es (S uden ’s - es ) and
an analysis o a iance (ANOVA) wi h a 95 % con idence le el (p <
0.05). This analysis was ca ied ou using he SPSS18 Excel s a is ical
package (Mic oso , USA).
3. Resul s and discussion
3.1. P epa a ion o bioplas ics
The i s s ep o he he mochemical p ocedu e o he p epa a ion o
bioplas ics ma ices is mixing, in which he di e en aw ma e ials we e
in oduced in o a mixe wi h di e en ein o cemen me hods. Fig. 1
shows he e olu ion o o que (M) and he p o iles o he pe cen age
a io be ween he inc ease in he ini ial empe a u e (100⋅(T-T
0
)/T
0
).
In his case, he p o iles a e simila wi hou showing signi ican
di e ences in he o que associa ed wi h he di e en blends. The eby,
in all cases, he p o ile shows a as inc ease a he beginning in o que,
ollowed by a dec ease ill a cons an alue, which is main ained du ing
he es o he p ocess. Thus, he ein o ced bioplas ics wi h gela in and a
pos -hea ea men show simila beha iou , dec easing o que while
he concen a ion o hese compounds o empe a u es inc eases in he
ma ix. Howe e , in he es o he sys ems, he e is no a b igh change,
highligh ing he sys ems Glyx-1, which possesses an appa en a ia ion,
maybe due o he enhancemen in he p ope ies wi h limi ed concen-
a ions o glyoxal. The sys em wi h a 20 w % o gela in (G-20) shows a
minimum o que, ollowed by a maximum o que in he sys em wi h a 1
w % o glyoxal (Glyx-1).
On he o he hand, all he he mal p o iles display a simila e olu-
ion, consis ing o a mode a e inc ease in empe a u e while he o que
alues inc ease. Thus, gene ally, he mal p o iles shown an exponen ial
shape whose a io is highe while mos o he concen a ion inc eases.
3.2. Cha ac e iza ion o samples
3.2.1. Physicochemical p ope ies
3.2.1.1. Fou ie ans o m in a ed spec oscopy (FTIR). Fig. 2 plo s he
esul s ob ained o he FTIR p o ile o he di e en bioplas ic samples
de eloped. Analysing he di e ences be ween he sys ems is possible by
de e mining he bonds comp ising each sample, as each bond abso bs a
di e en wa enumbe s. Thus, all he sys ems p esen ed a p o ile simila
o he e e ence sys em. The main peaks and hei wa enumbe a e
displayed in Table 2. Fi s ly, a band be ween 3500 and 3000 cm-1 (wi h
C.M. G anados-Ca e a e al.
Fu u e Foods 11 (2025) 100524
3
a maximum peak a app oxima ely 3266 cm-1) showed he s e ching o
NH bonds p esen in amides A and B and he s e ching o OH bonds
(Tü ke -Kaya and Huck, 2017). The 2928 and 2876 cm
−1
bands co e-
sponded o CH
2
s e ching (asymme ic and symme ic) and he 1620
cm
−1
peak co esponded o C =O s ain, all o hem p esen ed in he
p o ein chains (Tü ke -Kaya and Huck, 2017). None heless, he mos
ema kable bands o he p o eins a e hose ep esen ed in 1628, 1550
and 1228 cm-1, which ep esen ed amide I, II and III, espec i ely,
highligh ing hese amide bands which showed a maximum alue when a
hea ea men o 120 ◦C o 24 h was applied as a consequence o he
chemical modi ica ion ha su e ed he p o ein du ing he
manu ac u ing p ocess (Jim´
enez-Rosado e al., 2022). Finally, o he
bands shown a 1393 and 1100–974 cm
−1
a e ela ed o CH
2
and
C
–
O-C, espec i ely (Bake e al., 2014; Pe ez-Puyana, 2023).
3.2.2. Mechanical p ope ies
3.2.2.1. Dynamic lexu e es s. The lexu al p ope ies o he di e en
bioplas ics ob ained om he s ain sweep es s a e ep esen ed by he
c i ical s ain alues (Table 3). The s ain p o iles a e also shown in
Figu e S1. In all cases, he c i ical s ain alues a e signi ican ly lowe
han hose ob ained in he e e ence sys ems. The addi ion o gela in
wo sens he c i ical s ain (lowe alues) when i s p opo ion inc eases,
possibly due o he la ge di e ence in pa icle size o bo h p o eins (in
he case o SPI and gela in, hei pa icle size was 10 – 100 and 250 – 600
µm, espec i ely). The addi ion o saccha ose and ansglu aminase
(enzyma ic c osslinking) has a simila e ec bu wi hou signi ican
di e ences in he concen a ion used. Finally, he hea ea men and he
addi ion o glyoxal (physical and chemical c osslinking, espec i ely)
Fig. 1. E olu ion o mixing o que and pe cen age a io be ween he inc ease in empe a u e and he ini ial empe a u e (100⋅(T-T
0
)/T
0
) o SPI blends wi h di e en
ein o cemen me hods: (A) Gela in (5, 10 and 20 w % shown as G-5, G10 and G-20, espec i ely), (B) Saccha ose (5, 10 and 20 w %, shown as S-5, S-10 and S-20,
espec i ely), (C) The mal ea men (50, 120 ◦C shown HT-50 and HT-120, espec i ely), glyoxal (1 and 3 w %, shown as Glyx-1 and Glyx-3, espec i ely) and
ansglu aminase (0.1 and 0.2 w % shown as Tgase-0.1 and Tgase-0.2, espec i ely). Re . e e s o a e e ence sys em wi hou any ea men o addi i e included.
Fig. 2. FTIR p o iles o SPI blends wi h di e en ein o cemen me hods: (A) Gela in (5, 10 and 20 w % shown as G-5, G10 and G-20, espec i ely), (B) Saccha ose
(5, 10 and 20 w %, shown as S-5, S-10 and S-20, espec i ely), (C) The mal ea men (50, 120 ◦C shown HT-50 and HT-120, espec i ely), glyoxal (1 and 3 w %,
shown as Glyx-1 and Glyx-3, espec i ely) and ansglu aminase (0.1 and 0.2 w % shown as Tgase-0.1 and Tgase-0.2, espec i ely). Re . e e s o a e e ence sys em
wi hou any ea men o addi i e included.
Table 2
FTIR measu emen s o he di e en sys em de eloped.
Name in he
spec um
Co esponden peak (cm
−1
) Assigna ion
Fig. 2.AFig. 2.BFig. 2.C
A Amide A 3289 3266 3267 N-H s ain
B Amide B 2928 2934 2945 Asymme ic s ain CH
2
C CH
2
2852 2876 2876 Symme ic s e ching
CH
2
D Amide I 1620 1628 1628 C =O s ain
E Amide II 1529 1550 1520 N
–
H bending
F CH
2
1417 1393 1384 CH
2
olding
G Amide III 1215 1228 1237 N-H bending
H C
–
O
–
C 1106–964 1101–974 1111–974 C-O-C s e ching
Table 3
Pa ame e s ob ained om lexu e es s: C i ical s ain, E
′
and an δ a 1 Hz
(indica ed as E
′
1
and an (δ)
1
).
Sys em C i ical s ain (%) E’
1
(MPa) an (δ)
1
(-)
Re 1.000 ±0.05 820 ±140 0.27 ±0.02
G-5 0.700 ±0.90 120 ±3 0.29 ±0.01
G-10 0.600 ±0.30 231 ±19 0.27 ±0.01
G-20 0.200 ±0.05 216 ±88 0.39 ±0.01
S-5 0.315 ±0.01 1500 ±46 0.27 ±0.01
S-10 0.315 ±0.01 1370 ±134 0.28 ±0.01
S-20 0.315 ±0.01 1334 ±57 0.29 ±0.01
HT-50 0.780 ±0.01 251 ±88 0.28 ±0.02
HT-120 0.150 ±0.05 321 ±439 0.13 ±0.01
Glyx-1 0.320 ±0.01 2340 ±469 0.23 ±0.01
Glyx-3 0.170 ±0.20 2030 ±577 0.22 ±0.03
Tgase-0.1 0.310 ±0.01 2900 ±126 0.24 ±0.01
Tgase-0.2 0.310 ±0.01 2740 ±21 0.25 ±0.01
C.M. G anados-Ca e a e al.
Fu u e Foods 11 (2025) 100524
4
we e he ones ha a ec ed he c i ical s ain he mos , wi h he lowes
alues being achie ed. This beha iou may be due o a g ea e c oss-
linking be ween he chains ha s i ens he bioplas ics, lowe ing hei
elas ic condi ion (D¨
o s ein e al., 2018; Jim´
enez-Rosado e al., 2022).
F equency sweep es s a e plo ed in Fig. 3 in o de o assess he
s abili y o he bioplas ics. All he sys ems ha e a p edominan solid
cha ac e (E’ >E’’), showing ce ain ins abili y in hei mechanical
esis ance a high equencies (sho eco e y imes). I should be no ed
ha his slope is lowe in he case o chemical and physical c osslinking,
showing g ea e esis ance and being consis en wi h he alues ob-
ained o c i ical s ain. Table 3 also shows he alues o E’ and an (δ) a
1 Hz (E’
1
and an (δ)
1
) o imp o e he compa ison be ween he sys ems.
As can be seen, he inco po a ion o gela in in he bioplas ics p omo es a
dec ease in E’
1
and a g ea e dependence o iscoelas ic moduli on
equency. The esul s show ha he e is a sligh a ia ion in he
iscoelas ic p ope ies as a unc ion o he gela in concen a ion due o
he pa icle size, as p e iously men ioned. This educ ion in E’
1
was also
obse ed in he bioplas ic subjec ed o a hea ea men . On he o he
hand, he addi ion o saccha ose, glyoxal o ansglu aminase o he
samples causes an inc ease in E’
1
; howe e , as he amoun o saccha ose
addi i e in he bioplas ics inc eases, E’
1
dec eases, sugges ing ha hese
componen s imp o e he mechanical p ope ies by s eng hening he
s uc u e, bu only up o a ce ain poin . As o he loss angen , no
signi ican di e ences a e obse ed, excep o wo speci ic cases: he G-
20 sys em, which has he highes alue, showing he loss o pa o i s
solid cha ac e ; and he HT-120 sys em, which has he lowes alue,
being he mos igid sys em. These esul s a e consis en wi h he ones
ob ained in p e ious s udies whe e he inco po a ion o a pos - he mal
ea men p omo es a signi ican inc ease in E’ alues as well as a
dec ease in loss angen , o igina ing sys em mo e s able (Pe ez-Puyana,
2023). The inco po a ion o glyoxal can be ansla ed as a d as ic in-
c ease in E’ alues and he addi ion o ansglu aminase p omo es a
educ ion in he elonga ion o he sys ems (Cui e al., 2017; Pe -
ez-Puyana e al., 2022).
3.2.2.2. Tensile es s. The ensile p ope ies o he di e en bioplas ics
a e shown in Fig. 4. In all cases, he s ess-s ain cu es show an ini ial
linea elas ic egion whe e he e is a cons an slope be ween s ess and
s ain. La e , his linea egion is ollowed by a plas ic de o ma ion
phase, which shows a dec ease in he s ess-s ain slope when hese
bioplas ics exceed he elas ic limi , p omo ing he ac u e o he sample.
In pa icula , his beha iou is simila o he one demons a ed in p e-
ious s udies o soy p o ein-based bioplas ics by o he au ho s
(Fe n´
andez-Espada e al., 2016; Game o e al., 2019), exis ing a mini-
miza ion o he s ain a b eak and an enhancemen o s i ness wi h
mos o he ein o cemen me hods as shown in o he s udies (Boey
e al., 2022).
Table 4 shows he alues o Young’s modulus, maximum s ess and
s ain a b eak o he di e en sys ems. I is e iden ha he addi ion o
ein o cemen s educes he de o mabili y o he sys ems. This educ ion
is mo e p onounced when a biopolyme such as gela in o saccha ose is
added, possibly due o he lack o homogenisa ion be ween he bio-
polyme s as shown in o he s udies whe e he inco po a ion o ke a in
in o a SPI-bioplas ic, enhances he esis ance o he ma e ial bu , as a
esul o he he e ogenei y, he e is a educ ion in elonga ion (Wang,
2022). Fu he mo e, in he s udy de eloped by Om ani-Fa d e al., he
inco po a ion o gela in in o a whey p o ein bioplas ic modi ied he
p ope ies o he esul ing bioplas ics (Om ani-Fa d e al., 2020).
Howe e , he sys em wi h he lowes s ain a b eak is HT-120, likely
because he hea causes a dena u a ion o he soy p o ein, weakening he
bioplas ic s uc u e. This beha iou has been al eady desc ibed in p e-
ious s udies in which he e is a de e io a ion in he p ope ies as a esul
o high empe a u es (Baima k e al., 2021; Je ez e al., 2007). This
educ ion is linked o an inc ease in Young’s modulus o each sys em,
indica ing ha he ein o cemen leads o highe s i ness and lowe
de o mabili y. No ably, he HT-50 sys em is an excep ion, as i shows no
signi ican di e ences compa ed o he e e ence sys em and esul s in a
lowe Young’s modulus due o he lowe igidi y obse ed in dynamic
es s.
3.2.3. Func ional p ope ies
3.2.3.1. Wa e up ake capaci y and soluble ma e loss. Fi s ly, he wa e
con ac angle was measu ed o s udy he hyd ophilici y o hyd opho-
bici y o he di e en samples c ea ed. In his case, he measu emen s
we e eally di icul o do as a consequence o he exis ence o cons an
a ia ion in he alue due o he exis ence o high abso p ion. In all
cases, he wa e con ac angle was <57◦, which was he alue associa ed
wi h he e e ence sys ems and wi hou p esen ing signi ican di e -
ences be ween sys ems. These esul s indica e ha he bioplas ics p o-
duced ha e a mode a e hyd ophilic p ope y. This hyd ophilic
beha iou may be ad an ageous o applica ions whe e biodeg adabili y
and in e ac ion wi h he aqueous en i onmen a e mo e ele an such as
esh ood packaging (Audeb and e al., 2013), packaging o p oduc s in
low humidi y condi ions (Jin and Zhang, 2008) and coa ing applica ions
o ood o ag icul u al p oduc s (K och a and Mulde -Johns on, 1997).
Mo eo e , he measu emen s o wa e up ake capaci y and soluble
ma e loss a e ep esen ed in Fig. 5. Rein o cemen s gene ally educe
he wa e abso p ion capaci y o soy bioplas ics, which is less p o-
nounced when enzyma ic ( ansglu aminase) o physical (hea ea -
men ) c osslinking is inco po a ed. These esul s a e consis en wi h
wha has been obse ed o he mechanical p ope ies since a lowe
s i ness allows a g ea e deg ee o swelling, which allows mo e wa e o
be abso bed (´
Al a ez-Cas illo e al., 2018; Cuad i e al., 2016; Ku aishi
e al., 2001). Thus, in his case, he dec ease in he alues o wa e up-
ake capaci y is ela ed o he inc ease in he s i ness o he sys ems,
a oiding he inco po a ion o mo e wa e , and no due o he wa e
con ac angle p e iously men ioned because sys ems we e mo e
Fig. 3. F equency sweep es o SPI blends wi h di e en ein o cemen me hods: (A) Gela in (5, 10 and 20 w % shown as G-5, G10 and G-20, espec i ely), (B)
Saccha ose (5, 10 and 20 w %, shown as S-5, S-10 and S-20, espec i ely), (C) The mal ea men (50, 120 ◦C shown HT-50 and HT-120, espec i ely), glyoxal (1 and
3 w %, shown as Glyx-1 and Glyx-3, espec i ely) and ansglu aminase (0.1 and 0.2 w % shown as Tgase-0.1 and Tgase-0.2, espec i ely). Re . e e s o a e e ence
sys em wi hou any ea men o addi i e included.
C.M. G anados-Ca e a e al.
Fu u e Foods 11 (2025) 100524
5
hyd ophilic wi h he di e en ein o cemen me hods, being able o
abso b la ge amoun s o wa e . Thus, he inco po a ion o gela in in o a
bioplas ic ma ix p omo es a dec ease in wa e up ake capaci y, c ea ing
mo e hyd ophobic sys ems o wi h a wa e - epellen capaci y, as shown
in p e ious s udies (Chen e al., 2023). In he same way, in he case o
he addi ion o suga , an inc ease in suga con en can lead o a dec ease
in wa e up ake capaci y due o a educ ion in he hyd ophilici y as a
esul o he es ic ion in he mo emen o he polysaccha ide chains
(Halima ul e al., 2019).
As o SML, all sys ems ha e a simila beha iou , losing all he
plas icize and pa o he p o ein ha is solubilized. The HT-120 sys em
has he lowes loss o ma e ial, which is due o he loss o wa e no
conside ed du ing i s he mal ea men .
3.2.4. Biodeg adabili y
Fig. 6 depic s he biodeg ada ion o samples con aining gela in,
which is an essen ial ac o o e alua ing he e ec on he pollu ion o
he en i onmen and can be a ec ed by mul iple abio ic and bio ic
ac o s (Abe e al., 2024). Speci ically, he inclusion o hese bio-
polyme s in he ma ix leads o a signi ican inc ease in deg ada ion
speed as he gela in concen a ion ises. This may be due o he sys em’s
he e ogenei y, making i mo e suscep ible o mic obial a ack. Thus, as
desc ibed in p e ious s udies, he p esence o he e ogeneous su aces
enhances deg adabili y as a esul o a weak ne wo k o med (Liang
e al., 2022). The esul s show ha a e 20 days in bu ial condi ions,
hese bioplas ic ma e ials unde go se e e deg ada ion, esul ing in
educed hickness and weigh . Fu he mo e, he bioplas ic samples ha e
become signi ican ly mo e agile, making hem suscep ible o manual
b eakage.
4. Conclusion
Soy-p o ein-based bioplas ics we e success ully de eloped,
combining di e en ein o cemen me hods and demons a ing a
gene ally enhanced modi ica ion in he mechanical, physicochemical
and unc ional p ope ies in compa ison o he e e ence sys ems.
Pa icula ly, he addi ion o gela in and saccha ose imp o es he
mechanical p ope ies while dec easing he c i ical s ain and WUC.
The e o e, in he case o hea ea men , bioplas ics show a highe
s i ness and lowe de o mabili y, as well as a dec ease in WUC. Finally,
chemical and enzyma ic c osslinking ( he addi ion o glyoxal and
ansglu aminase, espec i ely) p omo es a simila ein o cemen ,
causing he highes Young’s modulus, speci ically app eciable when he
con es in he addi i e inc eases.
In addi ion, he inco po a ion o he di e en ein o cemen me hods
p omo es he o ma ion o bioplas ics wi h an inc eased s i ness and a
lowe de o mabili y, ob aining ein o ced ma e ials wi h be e me-
chanical p ope ies. Howe e , he ein o ced bioplas ics show a
lowe ing in he wa e up ake capaci y and he soluble ma e loss as a
esul o he minimisa ion in s i ness, as well as he inco po a ion o
hyd ophobic componen s in he ne wo ks. Thus, he sys ems wi h he
bes p ope ies we e G-5, S-10 and HT-120.
Fo his pu pose, u u e s udies should ocus on he e alua ion o he
he mal p ope ies o bioplas ic ma e ials and an analysis o hei
mic os uc u e by mic oscope echniques, as well as p o ing hei sui -
abili y as packaging ma e ial by, o ins ance, e alua ing he mig a ion
o he ma e ials in o ood simulan s. Mo eo e , in he case o he addi-
ion o p o ein as an addi i e, he e is a po en ial o op imisa ion in he
size o pa icles o u he enhance he eliabili y o hese bioplas ics.
E hical s a emen
Au ho s decla e ha he p esen esea ch does no in ol e any
human o animal s udy.
Au ho ’s name A ilia ion
Ca men Ma ía G anados-Ca e a Uni e si y o Se ille
Daniel Cas o-C iado Uni e si y o Se ille
Me cedes Jim´
enez-Rosado Uni e si y o Le´
on
Albe o Rome o Uni e si y o Se ille
Víc o Manuel Pe ez-Puyana Uni e si y o Se ille
CRediT au ho ship con ibu ion s a emen
Ca men Ma ía G anados-Ca e a: W i ing – o iginal d a ,
Fig. 4. S ess-s ain cu es o SPI blends wi h di e en ein o cemen me hods: (A) Gela in (5, 10 and 20 w % shown as G-5, G10 and G-20, espec i ely), (B)
Saccha ose (5, 10 and 20 w %, shown as S-5, S-10 and S-20, espec i ely), (C) The mal ea men (50, 120 ◦C shown HT-50 and HT-120, espec i ely), glyoxal (1 and
3 w %, shown as Glyx-1 and Glyx-3, espec i ely) and ansglu aminase (0.1 and 0.2 w % shown as Tgase-0.1 and Tgase-0.2, espec i ely). Re . e e s o a e e ence
sys em wi hou any ea men o addi i e included.
Table 4
Maximum ension, s ain a b eak and Young’s modulus o SPI blends wi h
di e en ein o cemen me hods.
Sys em Maximum
ension
(MPa)
S ain a b eak (mm/
mm)
Young’s Modulus
(MPa)
Re 1.10 ±0.20 1.09 ±0.12 27 ±2
G-5 1.43 ±0.24 0.13 ±0.08 38 ±7
G-10 1.88 ±0.28 0.20 ±0.06 53 ±11
G-20 1.70 ±0.18 0.23 ±0.05 35 ±7
S-5 1.20 ±0.20 0.16 ±0.03 31 ±7
S-10 1.08 ±0.05 0.16 ±0.03 33 ±2
S-20 0.45 ±0.06 0.07 ±0.01 24 ±6
HT-50 2.50 ±0.10 1.16 ±0.17 18 ±1
HT-120 6.00 ±1.40 0.02 ±0.01 343 ±53
Glyx-1 2.60 ±0.20 0.58 ±0.03 56 ±8
Glyx-3 3.50 ±0.30 0.54 ±0.04 45 ±6
Tgase-
0.1
1.80 ±0.10 0.34 ±0.03 53 ±2
Tgase-
0.2
1.80 ±0.30 0.24 ±0.03 54 ±9
C.M. G anados-Ca e a e al.
Fu u e Foods 11 (2025) 100524
6
Visualiza ion, So wa e, Me hodology, In es iga ion, Da a cu a ion.
Daniel Cas o-C iado: W i ing – o iginal d a , So wa e, In es iga ion,
Da a cu a ion, Concep ualiza ion. Me cedes Jim´
enez-Rosado: W i ing
– e iew & edi ing, Visualiza ion, Me hodology, Fo mal analysis,
Concep ualiza ion. Albe o Rome o: W i ing – e iew & edi ing, Su-
pe ision, Resou ces, Funding acquisi ion, Fo mal analysis. Víc o
Manuel Pe ez-Puyana: W i ing – e iew & edi ing, Valida ion,
Resou ces, P ojec adminis a ion.
Decla a ion o compe ing in e es
The au ho s decla e ha hey ha e no known compe ing inancial
in e es s o pe sonal ela ionships ha could ha e appea ed o in luence
he wo k epo ed in his pape .
Fig. 5. Wa e up ake capaci y (WUC) and soluble ma e loss (SML) o SPI blends wi h di e en ein o cemen me hods: (A) Gela in (5, 10 and 20 w % shown as G-5,
G10 and G-20, espec i ely), (B) Saccha ose (5, 10 and 20 w %, shown as S-5, S-10 and S-20, espec i ely), (C) The mal ea men (50, 120 ◦C shown HT-50 and HT-
120, espec i ely), glyoxal (1 and 3 w %, shown as Glyx-1 and Glyx-3, espec i ely) and ansglu aminase (0.1 and 0.2 w % shown as Tgase-0.1 and Tgase-0.2,
espec i ely). Re . e e s o a e e ence sys em wi hou any ea men o addi i e included.
Fig. 6. Biodeg adabili y o SPI/Gly bioplas ics combined wi h gela in.
C.M. G anados-Ca e a e al.
Fu u e Foods 11 (2025) 100524
7
Acknowledgmen s
Au ho s acknowledge he inancial suppo o he Spanish Go e n-
men (MICIU/AEI/10.13039/501100011033/ERDF/EU) h ough he
sponso ed p ojec wi h e . PID2021–124294OB-C21. In addi ion, his
esea ch has been co ounded by UE - Minis e io de Hacienda y Funci´
on
Pública - Fondos Eu opeos - Jun a de Andalucía - Conseje ía de Uni-
e sidad, In es igaci´
on e Inno aci´
on (SOL2024-31712). Au ho s would
also like o acknowledge CITIUS o g an ing access o he FTIR se ices.
Supplemen a y ma e ials
Supplemen a y ma e ial associa ed wi h his a icle can be ound, in
he online e sion, a doi:10.1016/j. u o.2024.100524.
Da a a ailabili y
Da a will be made a ailable on eques .
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