Ci a ion: Poˇ ízka, J.; Sla íko á, Z.;
Bidmono á, K.; Vymˇe alo á, M.;
Di iš, P. Physiochemical and Senso y
P ope ies o B ead Fo i ied wi h
Whea B an and Whey P o ein
Isola es. Foods 2023,12, 2635.
h ps://doi.o g/10.3390/
oods12132635
Academic Edi o : Lubomi Lapcik
Recei ed: 29 May 2023
Re ised: 27 June 2023
Accep ed: 1 July 2023
Published: 7 July 2023
Copy igh : © 2023 by he au ho s.
Licensee MDPI, Basel, Swi ze land.
This a icle is an open access a icle
dis ibu ed unde he e ms and
condi ions o he C ea i e Commons
A ibu ion (CC BY) license (h ps://
c ea i ecommons.o g/licenses/by/
4.0/).
oods
A icle
Physiochemical and Senso y P ope ies o B ead Fo i ied wi h
Whea B an and Whey P o ein Isola es
Ja omí Poˇ ízka 1,* , Zuzana Sla íko á1, Ka olína Bidmono á1, Mi osla a Vymˇe alo á2and Pa el Di iš 1
1Facul y o Chemis y, B no Uni e si y o Technology, 612 00 B no, Czech Republic;
[email p o ec ed] (Z.S.); [email p o ec ed] (P.D.)
2Mlýny J. Voženílek, L d., P ˚umyslo á107, 503 02 Pˇ edmˇeˇ ice nad Labem, Czech Republic;
[email p o ec ed]
*Co espondence: [email p o ec ed]
Abs ac :
This s udy in es iga ed he e ec o o i ying baked goods wi h whea b an (WBPI) and
whey p o ein isola es (WPI) on hei physicochemical and senso y p ope ies. The aim was o
enhance he nu i ional alue by inco po a ing high-p o ein ing edien s. WBPI and WPI, which a e
ich in essen ial amino acids, we e chosen o c ea e high-p o ein lou blends. The main ad an age
o WBPI is ha i is de i ed om eadily a ailable and inexpensi e whea b an. High-p o ein
lou blends o i ied wi h subs i u ions o 5%, 10%, and 15% lou wi h WBPI and WPI we e
subjec ed o chemical and heological analysis. WBPI subs i u ion sligh ly inc eased wa e binding
and so ening, bu i esul ed in a dec ease in dough quali y. In con as , WPI subs i u ion p olonged
dough de elopmen ime, imp o ed dough s abili y, and enhanced a inog aphic quali y. WBPI-
subs i u ed dough exhibi ed compa able ex ensog aphic p ope ies o he e e ence lou , wi h 5%
WBPI subs i u ion leading o imp o ed ene gy and dough esis ance. Howe e , as he le el o WBPI
lou subs i u ion inc eased, ex ensog aphic pa ame e s g adually declined wi hou u he enhancing
he dough’s mechanical p ope ies. Samples wi h 5% WPI subs i u ion demons a ed supe io
mechanical p ope ies compa ed o he e e ence sample. Bague e wi h high WBPI subs i u ion
was associa ed wi h educed o e all accep ance due o a bi e as e caused by he p esence o small
pep ides, e ulic acid, and annins, as con i med by co ela ion analysis.
Keywo ds:
high p o ein ood; pas y; whea b an; p o ein isola es; whey p o ein; heology;
senso y analysis
1. In oduc ion
Pas ies a e a popula ood wo ldwide and a e a egula pa o many people’s die s.
Cu en ly he e is g owing in e es in c ea ing pas ies wi h highe nu i ional alue and
ewe calo ies. This has led esea che s and ood manu ac u e s o explo e he po en ial
o o i ying pas ies wi h unc ional ing edien s ha can p o ide addi ional nu i ional
bene i s. Inc easing he p o ein con en by using p o ein isola es is cu en ly one o he
mos p omising app oaches o he p oduc ion o modi ied ood [1–7].
P o ein addi i es in gene al can be used as unc ional ing edien s in p epa ing high-
p o ein oods and can be o bo h animal and plan o igin. Physiochemical p ope ies ha
can be a ec ed by he addi ion o p o ein isola es, include ex u e, colo , and nu i ion. Fo
example, he addi ion o p o eins o pas y o mula ions can lead o changes in heology and
speci ic olume, which may esul in changes in he ex u e and mou h eel. Addi ionally,
he polyphenols p esen in plan isola es can cause he inished p oduc o ha e a da ke
colo . Plan p o ein isola es in pas ies can also inc ease he nu i ional alue by inc easing
p o ein and ibe while educing a [8–10].
One isola e wi h a high po en ial o use in he baking indus y is whea b an p o ein
isola e (WBPI). I is isola ed om he ou e laye o he whea g ain h ough a se ies o
Foods 2023,12, 2635. h ps://doi.o g/10.3390/ oods12132635 h ps://www.mdpi.com/jou nal/ oods
Foods 2023,12, 2635 2 o 15
p ocessing s eps ha include de a ing, solubiliza ion, and p ecipi a ion. The essen ial
ad an age o WBPI is ha i is p oduced om commonly a ailable whea b an, which is a
cheap and sus ainable ma e ial p oduced in la ge quan i y du ing milling [11].
WBPI is a plan -based p o ein isola e wi h a well-balanced amino acid p o ile ha is
compa able o plan p o eins like soy and pea and has a e y simila P o ein Diges ibili y
Co ec ed Amino Acid Sco e (PDCAAS). The isola e is pa icula ly ich in aspa agine,
glu amic acid, leucine, a ginine, and p oline. In addi ion o i s p o ein con en , WBPI also
con ains bioac i e compounds such as polyphenols, phy os e ols, and lignans ha ha e
been linked wi h heal h bene i s, such as imp o ed ca dio ascula heal h, an ioxidan
ac i i y, and an icance p ope ies [12].
The ood applica ion o WBPI has no been ex ensi ely s udied al hough a ew s udies
ha e desc ibed i s unc ional p ope ies in model sys ems. I has been p o en ha WBPI has
he po en ial o applica ion in ood o mula ion due o i s good emulsi ying and oaming
p ope ies and high wa e and a holding capaci y [
13
]. WBPI has also been expe imen ally
used as a p o ein o i ie in pas a [
14
] and b ead [
15
]. Spaghe i quali y was signi ican ly
a ec ed when 5–10% o WBPI was used o enhance he p o ein con en . The de e io a ion
o unc ional p ope ies was obse ed wi h highe WBPI subs i u ions, bu cooking loss
and appea ance we e s ill compa able o common whole-meal pas a. The e alua ion o he
in luence o WBPI o i ica ion on he unc ional p ope ies o b ead is mo e complica ed.
Alzuwaid e al. [
14
] and U am e al. [
15
] achie ed di e en esul s when assessing he
e ec o o i ica ion on loa olume. F om a de ailed s udy o he esul s, a subs an ial
imp o emen in olume was achie ed when only he albumin ac ion o WBPI was used.
Non- ac iona ed WBPI had he opposi e e ec .
Based on p e iously published indings, i is e iden ha he e ec o o i ying
pas ies wi h WBPI on physiochemical and senso y p ope ies is s ill no su icien ly un-
de s ood. The aim o his s udy is o expand he a ailable knowledge wi h new indings
by using con en ionally used analy ical echniques om he ield o dough analysis such
as a inog aphy and ex ensog aphy. A baking expe imen was also pa o he s udy.
High-p o ein bague es we e p epa ed and subjec ed o a comp ehensi e senso y analysis.
This s udy also includes a di ec compa ison o WBPI and whey p o ein isola e (WPI) as
common ep esen a i es o animal p o eins o explo e he ad an ages and disad an ages
o bo h unc ional ing edien s.
2. Ma e ials and Me hods
2.1. P o ein Isola es and Analysis
The WBPI was ob ained om whea b an (Mlýny J. Voženílek, Pˇ edmˇeˇ ice nad Labem,
Czech Republic) using he pH shi me hod wi h modi ica ions (Figu e 1). The p o ein
ac ion was ex ac ed using a NaOH (Lach-Ne , Ne a o ice, Czech Republic) solu ion
(pH 10.5
±
0.1) in a a io o 1:20 (whea b an/NaOH solu ion) o 2 h wi h con inuous
s i ing. The liquid p o ein ex ac was sepa a ed om dep o eined whea b an by cen-
i uga ion (8000 c , 15 min, oom empe a u e) and i s pH was adjus ed o 4.0
±
0.05
using 1 M ci ic acid (Lach-Ne , Ne a o ice, Czech Republic). The p ecipi a ed WBPI was
collec ed a e cen i uga ion (8000 pm, 15 min, oom empe a u e), lyophilized, and used
o u he expe imen s.
Foods 2023, 12, x FOR PEER REVIEW 3 o 16
Figu e 1. Isola ion o WBPI.
The WPI used o he expe imen s was a comme cial whey p o ein (Vilgain, B no,
Czech Republic).
The p o ein con en and amino acid p o ile o WBPI we e de e mined. The amoun
o ni ogen was analyzed using Eu o ec o EA3100, and he p o ein was ob ained by
mul iplying he ni ogen con en by he con e sion ac o 6.31, which is speci ic o whea -
b an p o eins. The amino acid p o ile was de e mined by HPLC. A 10 mg sample o
p o ein ma e ial was hyd olyzed in 3 mL o 6 M HCl o 12 h a 110 °C. The solu ion was
d ied and econs i u ed in 5 mL o wa e . HPLC analysis was pe o med acco ding o he
me hod desc ibed by Go issen e al. [16]. A single quad upole LC/MSD iQ Agilen 1260
(Agilen , San a Cla a, CA, USA) was used wi h Ad anceBio Amino Acid Analysis (AAA),
3.0 × 100 mm column (Agilen , USA).
2.2. P epa a ion and Cha ac e iza ion o Flou Samples
Plain high-glu en whea lou T530 (Mlýny J. Voženílek, Czech Republic) was
selec ed o he p epa a ion o high-p o ein lou blends and o all baking expe imen s.
Fo i ica ion o lou was pe o med by he di ec mass subs i u ion o pa o he lou
wi h ei he WBPI o WPI. Composi ion o lou mix u es wi h p o ein isola es is p esen ed
in Table 1.
Table 1. Composi ion o lou mix u es.
Flou Con en (g) Isola e Con en (g)
T530 92.5 0
5% WBPI 87.9 4.6
10% WBPI 83.3 9.3
15% WBPI 78.6 13.9
5% WPI 87.9 4.6
10% WPI 83.3 9.3
15% WPI 78.6 13.9
2.2.1. De e mina ion o We Glu en
Exac ly 10 g o he lou mix u es we e weighed in o a po celain bowl. A 2% sodium
chlo ide solu ion was added o he p o ein isola e in a olume o 5–5.5 mL. Using a
spa ula, a mode a ely s iff dough was p epa ed and hen kneaded well and shaped in o a
small ball. The dough co e ed by a wa ch glass and le o 30 min. A e wa ds, he dough
was washed unde a gen le s eam o wa e using a sie e. The washing was s opped when
wa e d ople s expelled om he we glu en we e anspa en . This measu emen was
pe o med on samples o lou , lou wi h WBPI subs i u ion (5, 10, and 15%), and lou
wi h whea p o ein subs i u ion (5, 10, and 15%).
The con en o we glu en, exp essed as a weigh pe cen age o he p oduc ’s d y
ma e , was calcula ed using he ollowing equa ion:
𝑋=𝑚∙10 ∙100
100 −𝑤

,
whe e m is he weigh o washed dough, and w1 is he wa e con en (%).
Figu e 1. Isola ion o WBPI.
The WPI used o he expe imen s was a comme cial whey p o ein (Vilgain, B no,
Czech Republic).
Foods 2023,12, 2635 3 o 15
The p o ein con en and amino acid p o ile o WBPI we e de e mined. The amoun
o ni ogen was analyzed using Eu o ec o EA3100, and he p o ein was ob ained by
mul iplying he ni ogen con en by he con e sion ac o 6.31, which is speci ic o whea -
b an p o eins. The amino acid p o ile was de e mined by HPLC. A 10 mg sample o
p o ein ma e ial was hyd olyzed in 3 mL o 6 M HCl o 12 h a 110
◦
C. The solu ion was
d ied and econs i u ed in 5 mL o wa e . HPLC analysis was pe o med acco ding o he
me hod desc ibed by Go issen e al. [
16
]. A single quad upole LC/MSD iQ Agilen 1260
(Agilen , San a Cla a, CA, USA) was used wi h Ad anceBio Amino Acid Analysis (AAA),
3.0 ×100 mm column (Agilen , USA).
2.2. P epa a ion and Cha ac e iza ion o Flou Samples
Plain high-glu en whea lou T530 (Mlýny J. Voženílek, Czech Republic) was selec ed
o he p epa a ion o high-p o ein lou blends and o all baking expe imen s. Fo i ica ion
o lou was pe o med by he di ec mass subs i u ion o pa o he lou wi h ei he WBPI
o WPI. Composi ion o lou mix u es wi h p o ein isola es is p esen ed in Table 1.
Table 1. Composi ion o lou mix u es.
Flou Con en (g) Isola e Con en (g)
T530 92.5 0
5% WBPI 87.9 4.6
10% WBPI 83.3 9.3
15% WBPI 78.6 13.9
5% WPI 87.9 4.6
10% WPI 83.3 9.3
15% WPI 78.6 13.9
2.2.1. De e mina ion o We Glu en
Exac ly 10 g o he lou mix u es we e weighed in o a po celain bowl. A 2% sodium
chlo ide solu ion was added o he p o ein isola e in a olume o 5–5.5 mL. Using a spa ula,
a mode a ely s i dough was p epa ed and hen kneaded well and shaped in o a small
ball. The dough co e ed by a wa ch glass and le o 30 min. A e wa ds, he dough was
washed unde a gen le s eam o wa e using a sie e. The washing was s opped when
wa e d ople s expelled om he we glu en we e anspa en . This measu emen was
pe o med on samples o lou , lou wi h WBPI subs i u ion (5, 10, and 15%), and lou
wi h whea p o ein subs i u ion (5, 10, and 15%).
The con en o we glu en, exp essed as a weigh pe cen age o he p oduc ’s d y
ma e , was calcula ed using he ollowing equa ion:
X=m·10·100
100 −w1
,
whe e mis he weigh o washed dough, and w1is he wa e con en (%).
2.2.2. De e mina ion o Sedimen a ion Value
Exac ly 80 mL o lac ic acid was ans e ed in o a sedimen a ion lask, which was
hen b ough o a empe a u e o 27
◦
C. F om eshly washed glu en, 1 g was weighed and
o n in o app oxima ely 30 equal pieces. In a po celain dish, 10 mL o empe ed lac ic acid
was added along wi h he small pieces o glu en. The con en o he po celain dish was
ans e ed o he swelling lask con aining he main po ion o he lac ic acid solu ion. The
lask was hea ed in a wa e ba h o 27
◦
C and le o 120 min. A e 10 and 20 min, he
con en s o he lask we e gen ly mixed o p e en he glu en pieces om s icking oge he
o adhe ing o he bo om. The lask was s oppe ed in such a way ha he s oppe eached
he ze o ma k on he scale. By g adually il ing he lask, he swollen glu en pa icles slid
in o he calib a ed neck. The olume was immedia ely ead on he scale o he nea es
0.5 di ision. This measu emen was pe o med on glu en om T530 lou , glu en wi h
Foods 2023,12, 2635 4 o 15
WBPI subs i u ion (5, 10, and 15%) and glu en om lou wi h whey p o ein subs i u ion (5,
10, and 15%).
2.2.3. Wa e and Elemen al Analysis o Flou Samples
The wa e con en in all lou samples was analyzed using he Ka l Fishe me hod
de eloped by Lau a Co pa¸s e al. (2013) [
17
]. Ka l Fishe i a ion was pe o med on KF
Ti ando (Me ohm, He isau, Swi ze land). Elemen al analysis o C, H, N, S was pe o med
by he elemen al analyze Eu o ec o EA3100 (Eu o ec o , Pa ia, I aly).
2.2.4. Fa inog aphic Analysis o Flou Samples
The a inog aphic analysis was ca ied ou a he milling company Mlýny J. Voženílek
in Pˇ edmˇeˇ ice nad Labem (Czech Republic) on Fa inog aph-TS ins umen (B abende ,
Duisbu g, Ge many). The main componen o he a inog aph was a mixe wi h a capaci y
o 300 g equipped wi h blades o a ing a a speed o 63 e olu ions pe minu e. I also had
a he mos a ha ensu ed a cons an empe a u e o 30
◦
C du ing he expe imen . Exac ly
300 g o lou we e added o he mixe , and an app op ia e wa e olume was in oduced
using a bu e e. The wa e was added un il he o que alue eached 500 a inog aphic
uni s (FUs), which is s anda d o lou es ing in indus ial milling. Fa inog aphic analysis
was pe o med on samples o T530 lou , T530 + 5% WBPI, and T530 + 5% WPI. Highe
pe cen age concen a ions o WPI and WBPI we e no es ed because he analy ical limi o
500 FUs could no be eached wi h his ype o ins umen . Fa inog aphic indica o s we e
calcula ed:
Wa e abso p ion =
mwa e
m lou
×100
The dough-de elopmen ime indica ed he du a ion equi ed o dough o each
500 FUs. The s abili y o dough e e s o i s abili y o main ain i s s uc u e and esis
weakening o b eakdown du ing mixing while he o que emains a 500 FUs. The deg ee
o so ening indica ed a dec ease in dough esis ance o mixing ep esen ed as an FU a e
10 and 12 min. The a inog aphic quali y numbe (FQN) indica ed he ime om he
beginning o measu emen o he 30 FU d op om he maximum cu e.
2.2.5. Ex ensog aphic Analysis o Flou Samples
The ex ensog aphic analysis was pe o med on a Ex ensog aph-E ins umen (B aben-
de , Ge many) hea ed o 30
◦
C. The ins umen se ings a e lis ed in Table 2. The ex en-
sog aphic measu emen was conduc ed on T530 lou , T530 lou wi h 5, 10, and 15%
subs i u ion o lou wi h WBPI and WPI.
Table 2. Ex ensog aph se ings.
Speed o balling uni (min−1)83.0 ±3.0
Speed o dough oll (min−1)15.0 ±1.0
Speed o s e ching hook (mm·s−1)14.5 ±0.5
Ex ensog aphic o ce (mN·EJ−1)12.3 ±0.3
Dough was p epa ed using a mixe , which was pa o he a inog aph. Addi ionally,
6 g o sodium chlo ide we e weighed and dissol ed in he app op ia e amoun o wa e o
p epa e 0.1 M solu ion. This solu ion was used o p epa e he dough o 400 FU consis ency.
The op imized amoun o wa e o each mix u e is p o ided in Table 3.
The dough was o med in o a small ball using a shape and hen placed in o a shee e
and eshaped in o a cylinde . Dough cylinde s we e placed in o a chambe hea ed o 30
◦
C
and le o es o 30 min. A e he es ing pe iod, he dough was s e ched by he de ice’s
hook un il i b oke a e which i was kneaded again, eshaped in he shee e , and allowed
o es o ano he 30 min. Expe imen was epea ed. The esis ance exe ed by he hook on
he dough du ing s e ching was eco ded.
Foods 2023,12, 2635 5 o 15
Table 3. Op imized wa e addi ion o ex ensog aphic measu emen s.
Wa e Volume (mL)
T530 54.6
5% WBPI 52.6
10% WBPI 54.9
15% WBPI 57.7
5% WPI 47.9
2.2.6. Baking Expe imen s and Senso y Analysis o Pas y
Dough o he baking expe imen was p epa ed by mixing o lou blends, sal , yeas s
and wa e . The a io o he aw ma e ials is desc ibed in Table 4. The pas y was shaped
in o bague es and baked a 240
◦
C o 20 min. Senso y analysis made om e e ence and
o i ied lou was assessed by 20 e alua o s in a specialized labo a o y. Tex u al and la o
pa ame e s we e also e alua ed.
Table 4. Raw ma e ial o pas y p oduc ion.
Wa e (g) Flou (g) Isola e (g) Yeas (g) Sal (g)
T530
64
92.5 0
1.2 2.4
5% WBPI 87.9 4.6
10% WBPI 83.3 9.3
15% WBPI 78.6 13.9
5% WPI 87.9 4.6
10% WPI 83.3 9.3
15% WPI 78.6 13.9
2.2.7. S a is ical Analysis
Hypo hesis es ing was used o e alua e he impac o WBPI- and WPI- o i ied lou
on he physiochemical and senso y p ope ies o dough and pas y. The da ase was
p ocessed using he S a is ica so wa e (TIBCO, Palo Al o, CA, USA, e sion 13.0). ANOVA
and K uskal–Wallis ANOVA was used o assess s a is ically signi ican di e ences among
he o i ied bake y p oduc s. Hypo hesis es ing was conduc ed a a signi icance le el o
α= 0.05.
3. Resul s
3.1. Analysis o WBPI and WPI
An analysis o WBPI and WPI e ealed in e es ing amino acid p o iles, which a e
p esen ed in Table 5along wi h he p o ein con en . Bo h p o ein isola es a e ich in amino
acids ha a e impo an o spo s nu i ion. WPI is pa icula ly ich in leucine, lysine,
isoleucine, and aline. These b anched-chain amino acids (BCAAs) make up app oxima ely
70% o he amino acids in muscles. BCAAs a e ecognized o hei ole in p omo ing
muscle g ow h and acili a ing eco e y. P e ious s udies ha e shown ha hey can
inc ease he a e o p o ein syn hesis while educing p o ein deg ada ion du ing pe iods
o es . Consequen ly, a hle es commonly use hese amino acids as die a y supplemen s,
which a e o en inco po a ed in o mix u es e e ed o as BCAAs. As a esul , hese p oduc s
ha e gained popula i y in he ange o die a y supplemen s a ailable o a hle es. WPI
also con ains signi ican amoun s o h eonine, and among he non-essen ial amino acids,
glu amine and aspa ic acid a e he mos abundan [18].

Foods 2023,12, 2635 6 o 15
Table 5. P o ein con en and amino acid composi ion o p o ein isola es.
Amino Acid Con en
(%) WBPI WPI
Me 1.2 1.6
Me 1.2 1.6
Lys 2.7 6.8
Th 2.2 5.4
Asp 6.2 8.5
Se 3.2 4.9
Glu 13.6 13.7
Gly 3.5 1.5
Ala 3.1 3.8
Ty 2.5 2.2
Val 3.1 4.6
Phe 2.8 2.5
Ile 1.9 4.8
Leu 4.6 8.1
His 2.2 1.3
A g 5.2 1.9
Cys 1.1 1.6
P o 6.4 4.9
T p 0.6 1.3
P o ein con en (%) 67.6 76.9
WBPI also con ains a ange o nu i ionally impo an amino acids, and like whey
p o ein i has a signi ican amoun o essen ial BCAAs. The mos abundan non-essen ial
amino acid in WBPI is aspa ic acid, ollowed by glu amine and a ginine, which plays a ole
in egula ing u ea syn hesis in he li e and s imula ing he sec e ion o g ow h ho mone
and insulin. G ow h ho mone is impo an in he pos -physical pe o mance eco e y
phase [19].
Du ing s o age, he lysine in whea lou deg ades. P oduc s made om aw ma e ials
s o ed o a longe pe iod may ha e a lowe concen a ion o lysine. This de iciency can be
sol ed by subs i u ing a po ion o he lou wi h a p o ein isola e. Since bo h WBPI and
WPI a e ich in lysine, o i ying bake y p oduc s could supplemen his limi ing amino
acid [19].
WPI is known o i s high PDCAAS sco e o 1, whe eas WBPI, a plan -based p o ein,
has a lowe sco e o 0.54 wi h phenylalanine being he limi ing amino acid [20].
3.2. Analysis o Flou Samples
The p o ein, wa e , and we glu en con en and he glu en sedimen a ion olume o
he e e ence lou and he high-p o ein blends om WPI and WBPI subs i u ion we e
de e mined. All samples we e p epa ed ollowing he p ocedu es desc ibed in Sec ion 2.2.
The esul s o he analysis a e p esen ed in Table 6.
Table 6. Cha ac e is ics o lou and high p o ein blends.
P o ein Con en
(%)
Wa e Con en
(%)
We Glu en
Con en (%)
Sedimen a ion
Volume (mL)
T530 13.8 10.2 38.6 21.0
5% WBPI 16.5 9.8 33.5 25.0
10% WBPI 19.2 9.4 31.2 28.0
15% WBPI 21.9 9.1 28.3 42.0
5% WPI 16.9 10.0 33.1 23.0
10% WPI 20.1 9.8 30.5 20.0
15% WPI 23.3 9.7 19.6 17.0
Foods 2023,12, 2635 7 o 15
3.3. Rheological P ope ies o P epa ed Doughs
3.3.1. Fa inog aphic Analysis o Dough
The pu pose o he a inog aphic measu emen was o assess he impac o WBPI
and WPI subs i u ion on he heological p ope ies o he dough ollowing he p ocedu e
desc ibed in Sec ion 2.2.4. Table 7displays he alues o indi idual a inog aphic indica o s
o all samples: wa e abso p ion, de elopmen ime, s abili y, so ening deg ee, and
a inog aphic numbe . Two so ening deg ees we e eco ded: 10 min as measu ed om he
s a o he measu emen and 12 min as measu ed a e eaching maximum o que.
Table 7. Fa inog aphic cha ac e is ics o lou mix u es.
T530 5% WBPI 5% WPI
Wa e abso p ion [%] 55.8 56.4 52.6
De elopmen ime [min] 1.8 1.7 6.8
S abili y [min] 3.8 2.6 9.8
So ening deg ee (10) [FU] 72.0 73.0 20.0
So ening deg ee (12) [FU] 94.0 75.0 74.0
Fa inog aphic quali y numbe 35.0 30.0 113.0
A e e ence sample o T530 lou wi hou any p o ein subs i u ion was used. One
impo an physical cha ac e is ic o whea lou is i s wa e abso p ion capaci y. S ong
lou s ypically exhibi a wa e abso p ion ange o 55–60%, meaning ha 100 g o whea
lou can abso b 55 o 60 g o wa e [
21
]. The analyzed T530 lou ell wi hin his ange,
indica ing i s s eng h and sui abili y o b ead baking due o i s high glu en con en . Dough
de elopmen ime and dough s abili y indica e lou s eng h. The highe alues indica e a
s onge dough whe e whea glu en plays a c ucial ole in o ming a h ee-dimensional
iscoelas ic s uc u e. In he Czech Republic he pa ame e o dough s abili y ypically
anges om 3 o 5 min o whea lou s. The e e ence T530 lou , based on he measu emen ,
achie ed a s abili y alue o 3.8 min.
Da a om he a inog aph (Table 8) demons a ed ha eplacing whea lou wi h
WBPI esul ed in a sligh inc ease in wa e binding compa ed o he e e ence sample. This
inc ease could be a ibu ed o he highe p o ein con en and he p esence o ibe , which
con ains nume ous hyd oxyl g oups ha in e ac wi h hyd ogen bonds in wa e , he eby
enhancing abso p ion. This inding is consis en wi h he esul s o a s udy conduc ed by
Manuel Gómez e al. [
22
] ha in es iga ed he e ec o ex uded whea b an on dough
heology and b ead quali y. Subs i u ing lou wi h WBPI did no signi ican ly al e he
dough de elopmen ime: only by 0.1 min compa ed o he e e ence sample. Du ing dough
de elopmen , in e ac ions be ween ibe and glu en can occu , po en ially p olonging he
p ocess. Howe e , he WBPI, despi e i s ibe con en , did no lead o his phenomenon [
22
].
On he o he hand, dough s abili y exhibi ed a no iceable dec ease. The subs i u ion
o 5% WBPI esul ed in a weakening o he glu en ne wo k and a dec ease in lou s eng h.
Consequen ly, s abili y dec eased by 1.2 min compa ed o he e e ence sample, a signi ican
di e ence. Addi ionally, WBPI subs i u ion led o a slowdown in dough so ening, which
is in e sely ela ed o he a inog aph quali y numbe (FQN).
Fa inog aphic analysis was also conduc ed on lou blends wi h WPI. The da a om
he a inog aph (Table 8) indica ed a dec ease in wa e abso p ion compa ed o he e e ence
sample. Subs i u ing lou wi h WPI esul ed in he o ma ion o a complex sys em ha
hinde ed hyd a ion, ex ensibili y, and whea glu en alignmen , leading o an inc ease in
dough de elopmen ime by 5 min wi h 5% WPI. In addi ion, dough s abili y inc eased
by 6 min compa ed o he e e ence sample. These indings align wi h a s udy conduc ed
by Ind ani e al. [
23
], which demons a ed ha he subs i u ion o up o 10% whey p o ein
enhances dough s abili y. Howe e , a di e en conclusion was eached in he s udy
published by Tang e al. [
24
], who ound ha he subs i u ion o WPI caused a dec ease ha
was belie ed o be caused by in e ac ions be ween he sul hyd yl g oup in WPI and he
disul ide bond in whea glu en. In hei s udy, he subs i u ion o 10% WPI o he dough
Foods 2023,12, 2635 8 o 15
esul ed in a s abili y dec ease: s abili y ime alue, 6.5 min. These con as ing esul s
highligh ed he ac ha he subs i u ion o WPI o di e en ypes o lou had a ied e ec s
on heological p ope ies, sugges ing a need o u he s udy.
The so ening deg ee, which is de e mined by he di e ence be ween he consis ency
alue a he maximum and he consis ency alue a 10 min, dec eased due o longe dough
de elopmen ime. This implied ha as he de elopmen ime app oached 10 min, he
ime o he cu e o d op became sho e . I can be assumed ha he so ening ime a e
12 min will inc ease wi h a highe amoun o whey p o ein. Acco ding o he FQN, which
inc eased app oxima ely h ee old compa ed o he e e ence sample, he dough quali y
imp o ed wi h a 5% whey p o ein subs i u ion.
3.3.2. Ex ensog aphic Analysis o Dough
The aim o he ex ensog aph measu emen was o de e mine he in luence o he WBPI
and WPI subs i u ion on he heological p ope ies o he dough, acco ding o he p ocedu e
desc ibed in Sec ion 2.2.5. The alues o indi idual ex ensog aphic indica o s—ene gy,
esis ance, ex ensibili y, and a io numbe —-a e lis ed in Table 8. Two a io numbe s we e
eco ded du ing he measu emen . The a io numbe was be ween esis ance and ene gy,
and he a io numbe (max) was de ined as he a io be ween he maximum and ene gy.
P oblems wi h ex ensi e adhesi eness o he dough caused poo o ma ion o ille and
olle o he 10 and 15% o WPI subs i u ions. The dough s uck o he cylinde and olling
pin, making i impossible o comple e he ex ensog aph de e mina ion. The adhesi eness
o he dough could po en ially ha e been elimina ed by educing he wa e con en , bu o
bo h a inog aphic and ex ensog aphic de e mina ion, i was always necessa y o c ea e a
dough o maximum consis ency, eaching app oxima ely 500 FUs. Wi hin he ex ensog aph
de e mina ion, only he sample wi h 5% WPI subs i u ion was analyzed.
Table 8. Ex ensog aphic pa ame e s o T530 lou and high p o ein blends.
T530 5% WBPI 10% WBPI 15% WBPI 5% WPI
Res ing ime (min) 30 60 30 60 30 60 30 60 30 60
Ene gy (cm2)82 82 81 97 56 55 44 42 119 164
Resis ance (FU) 216 230 728 960 524 276 144 64 289 196
Duc ili y (mm) 185 176 85 80 58 54 53 51 196 188
Max (FU) 317 343 741 961 799 865 680 691 443 657
Ra io numbe 1.2 1.3 8.6 12.0 9.0 4.6 2.7 2.1 1.5 2.2
Ra io numbe (max) 1.7 2.0 8.8 12.0 13.9 16.1 12.9 13.6 2.3 3.5
The ene gy o he dough was conside ed a measu e o i s bake y wo kabili y and
dough quali y. The lowe he ene gy, he less esis an and s able he dough du ing
p ocessing. Flou s wi h a igid and sho glu en s uc u e ypically ha e low dough ene gy.
The measu ed ene gy alue o he e e ence sample was 82 cm
2
, which co esponded o
he s anda d o Czech lou s, whe e his pa ame e is a ound 90 cm2[25].
Subs i u ing lou wi h WBPI wi hin 30 min o dough de elopmen did no lead o
signi ican changes in ex ensog aphic pa ame e s compa ed o he e e ence lou . Howe e ,
a e 60 min om dough de elopmen , he ene gy inc eased by 16 cm
2
. This indica ed ha
he leng h o he in e al had an impac on he amoun o ene gy when subs i u ing up
o 5% WBPI. As he subs i u ion inc eased, he ene gy showed a dec easing end. The
dough wi h 15% WBPI subs i u ion was e alua ed as ha ing he lowes ene gy esis ance
and s abili y, which was hal ha o he e e ence sample.
Dough esis ance indica es he s eng h o glu en, meaning ha highe esis ance
esul s in i me glu en and a s onge and mechanically mo e esis an dough. The subs i-
u ion wi h 5% WBPI showed a esis ance app oxima ely ou imes highe han ha o he
e e ence sample. Simila o he ene gy pa ame e , esis ance also exhibi ed a dec easing
Foods 2023,12, 2635 9 o 15
end wi h inc easing WBPI subs i u ion. The lowes glu en s eng h was obse ed in he
case o 15% WBPI. The esul ing dough was s i bu less mechanically esis an .
Dough ex ensibili y indica ed he elas ici y o he dough. The e e ence sample showed
ex ensibili y a ound 180 mm, which a ied depending on he ma u a ion ime. The op imal
ex ensibili y o unmodi ied lou s used in baking anges om 140 o 170 mm [
2
]. The
sample wi h 5% WBPI subs i u ion exhibi ed app oxima ely hal he ex ensibili y compa ed
o he e e ence sample. Samples wi h highe WBPI subs i u ion we e cha ac e ized as mo e
b i le and less elas ic. This phenomenon can be connec ed o he p esence o die a y ibe
in WBPI because i in luences wa e edis ibu ion in glu en and causes physical damage o
he glu en ma ix, which can esul in educed glu en isco-elas ici y [26].
F om he pe spec i e o measu ing dough ene gy, esis ance, and ex ensibili y, he
dough p ope ies imp o ed wi h 5% WBPI subs i u ion, whe e bo h ene gy and esis ance
we e highe compa ed o he e e ence sample. This sugges s ha dough subs i u ed
wi h 5% WBPI exhibi s g ea e mechanical esis ance han he e e ence lou . On he
o he hand, samples wi h highe subs i u ion le els (10% and 15%) showed de e io a ing
mechanical esis ance.
In addi ion o esis ance, he maximum o he ex ensog aphic cu e also cha ac e ized
he s eng h o glu en. Simila o he esis ance pa ame e , he maximum pa ame e was
he highes in he sample wi h 5% WBPI. A signi ican dec ease in he maximum pa ame e
occu ed in he sample wi h 15% WBPI subs i u ion. This dec ease was wice as g ea as
ha o he e e ence lou (T530).
The a io numbe indica ed he a io be ween esis ance and ex ensibili y, unlike he
a io numbe (max), which indica ed he a io be ween he maximum and ex ensibili y.
The mos simila a io numbe o he e e ence sample was ound o he sample wi h 15%
WBPI subs i u ion. A e 60 min o ma u a ion, he samples showed a dec easing end,
excep o he 5% WBPI subs i u ion. The a io numbe (max) was he lowes in he 15%
WBPI sample, hus pu ing i he closes o he e e ence sample. The op imal alue o he
a io numbe should be 2–2.5 al hough i could be highe o special lou s. In he Czech
Republic, he a e age ex ensog aphic a io alls sligh ly below 2 o lou s o medium
quali y [
25
]. F om he o e all assessmen o he ex ensog aphic pa ame e s o he p epa ed
blends, i is e iden ha he dough wi h a 5% WBPI subs i u ion appea ed o be he bes
choice o u he p ocessing. As WBPI subs i u ion inc eased, he ex ensog aphic quali y
dec eased signi ican ly.
Mix u es wi h whey p o ein had signi ican ly di e en p ope ies compa ed o he
e e ence and mix u es wi h WBPI subs i u ion. Subs i u ing WPI lou led o a signi ican
inc ease in dough ene gy compa ed o he e e ence sample. The inc ease in dough ene gy
s eng hened wi h inc easing ime in e al om dough de elopmen . The high dough en-
e gy alue wi h 5% WPI subs i u ion indica es ha he dough is no sensi i e o p ocessing
condi ions and will no so en apidly du ing ma u a ion and p oo ing.
The dough esis ance alue o he mix u e wi h 5% WPI did no show he same
posi i e end as he ene gy men ioned ea lie in he case o WBPI subs i u ion. The
ex ensibili y o he dough wi h 5% WPI did no show a signi ican di e ence compa ed o
he e e ence sample.
Fo ene gy, esis ance, and ex ensibili y measu emen s, he dough p ope ies imp o ed
wi h 5% WPI, as bo h ene gy and ex ensibili y we e highe compa ed o he e e ence sam-
ple. The esis ance alue was no signi ican ly lowe han ha o he e e ence, indica ing
ha he dough o i ied wi h 5% WPI exhibi s simila mechanical s eng h o he e e -
ence sample.
The maximum dough alue wi h 5% WPI showed he highes alue a e a 60 min
es ing pe iod compa ed o he e e ence. Howe e , i was signi ican ly lowe han he
maximum alues o he 5% WBPI sample. The a io numbe did no e eal a signi ican
di e ence be ween i and he e e ence.