Impac o yeas and ungi (1→3)(1→6)-β-glucan concen a es on
iscoelas ic beha io and b ead making pe o mance o glu en- ee
ice-based doughs.
Sand a Pe ez-Qui ce, Ped o A. Caballe o, An onio Vela, Ma ina Villanue a, Felicidad
Ronda*
Depa men o Ag icul u e and Fo es y Enginee ing, Food Technology, College o Ag icul u al
and Fo es y Enginee ing, Uni e si y o Valladolid, A . Mad id, 44, 34004 Palencia, Spain
Co esponding au ho : onda@ia .u a.es
Abs ac
The addi ion o bioac i e β-glucan o glu en- ee b eads is o special in e es o people su e ing
om celiac disease. Mos o he s udies ound in li e a u e in ol e ce eal (1 → 3)(1 → 4)-β-
glucan, while hose om yeas and ungi a e s ill nea ly unexplo ed. This s udy ocuses on he
e ec o o i ying glu en- ee ice-based doughs and b eads wi h (1 → 3)(1 → 6)-β-glucan
concen a es de i ed om yeas s –soluble (SBG) and insoluble (IBG)– and ungi (Pleu o us
Os ea us) (FBG). SBG-en iched doughs we e less i m and exhibi ed lowe esis ance o
de o ma ion han doughs wi h FBG o IBG. In con as , FBG- and IBG-en iched doughs
inc eased hei esis ance o de o ma ion as he concen a ion inc eased. Doughs wi h a i me
consis ency as de e mined by a o wa d ex usion es (FBG- and IBG-en iched doughs)
co esponded o hose wi h la ge dynamic moduli and lowe equency dependence, lowe
elas ic de o ma ion and highe iscosi y a s eady s a e. The physical quali y o b eads was
signi ican ly imp o ed by addi ion o all ypes o (1→3)(1→6)-β-glucan a op imized dough
hyd a ion. They caused an inc ease in he speci ic olume o he b eads, a educ ion in hei
ha dness and longe shel -li e. Senso y e alua ion also demons a ed an imp o emen in b ead
o ganolep ic a ibu es when SBG was added.
Keywo ds: (1 → 3)(1 → 6)-β-glucan, heology, glu en- ee, dough, b ead
1. In oduc ion
β-glucan (BG) is a homoglucose polyme widely dis ibu ed in he cell walls o
mic oo ganisms, pa icula ly o he bake 's and b ewe 's yeas Saccha omyces ce e isiae,
mush ooms, and ce eals (mainly oa s and ba ley) (Ki isuban e al., 2014; Liu e al., 2008). The
di e ences in he mac omolecula s uc u e be ween β-glucans om di e en sou ces (yeas ,
ungal, and ce eal β-glucans) ha e been p e iously desc ibed by Volman e al. (2008). The β-
glucans o yeas and ungi consis o 1,3 β-linked glycopy anosyl esidues wi h small numbe s
o 1,6 β-linked b anches. In con as , oa and ba ley cell walls con ain unb anched β-glucans
wi h 1,3 and 1,4 β-linked glycopy anosyl esidues. BG om ce eals is classi ied as die a y ibe
ha is no hyd olyzed in he human diges i e ac , and is a non-calo i ic ing edien (Bo chani e
al., 2016). BG ob ained om yeas and mush ooms p esen s posi i e e ec s on human heal h,
such as immune-s imula ion, an i-in lamma o y, an imic obial, and an i- umo al esponses
(Chan e al., 2009; Kim e al., 2011; Rop e al., 2009; San ipanichwong & Suphan ha ika, 2009;
Wo asinchai e al., 2006). (1→3)(1→6)-β-glucans ha e also demons a ed hypoglycemic
ac i i y in animal s udies, and a e likewise known o hei hypocholes e olemic ac i i y and in
educing a he oscle osis (Mi anda-Nan es e al. 2011). Ce ain p epa a ions o yeas -de i ed
(1→3)(1→6)-β-glucan ha e ecen ly been app o ed as no el ood ing edien s by he Eu opean
Food Sa e y Au ho i y (EFSA) and gi en “Gene ally Recognized as Sa e” s a us by US Food
and D ug Adminis a ion. So a , he EFSA has no ye app o ed a heal h claim on immune
unc ion o yeas (1→3)(1→6)-β-glucan p epa a ions (EFSA, 2011; Samuelsen e al., 2014).
The cell wall o yeas (Saccha omyces ce e isiae) con ains abou 50-65% o (1→3)(1→6)-β-
glucan (Kogan & Koche , 2007). The di e ence in molecula and s uc u al ea u es acco ding
o he o igin o BG leads o di e ences in hei physical p ope ies and, he eby, di e en e ec s
on he unc ionali y o ood sys ems (Bancha hanakij & Suphan ha ika, 2009). The po en ial
applica ions o (1 → 3)(1 → 6)-β-glucan in ood s u s ha e p e iously been epo ed. This
includes as a hickening, wa e -holding, and oil-binding agen and emulsion s abilize , as a a
eplace in ood emulsions (San ipanichwong & Suphan ha ika, 2009; Wo asinchai e al.,
2006), and as a ex u e modi ie in s a ch gels. (Sa apai & Suphan ha ika, 2007). Howe e , i s
applica ion in en ichmen o glu en- ee p oduc s is mos ly unexplo ed, e en hough i holds
special in e es o people su e ing om celiac disease. These indi iduals ha e a signi ican a e
o o he associa ed ch onic diseases, such as gas i is, i amin B de iciency, ca diomyopa hy,
and skin p oblems, as well as obesi y-me abolic synd ome and diabe es, due o hei highe a
and calo ie-dense die s compa ed o he gene al popula ion (Le ne & Ma hias, 2015; Lebwohl
e al., 2014; Hele e al., 2009; C onin & Shanahan, 1997). Gi en he ecognized heal h bene i s
o (1→3)(1→6)-β-glucans, o i ying high-consump ion oods such as b ead is o g ea
impo ance o he gene al popula ion bu , in pa icula , o pa ien s wi h celiac disease
(Mi anda-Nan es e al. 2011). Howe e , only a ew s udies ha e ocused on (1 → 3)(1 → 6)-β-
glucan en ichmen o cakes (Kim e al., 2011), glu en- ee ice noodles (Heo e al., 2014), and
s a ch b eads (Ki isuban e al., 2014). Kim e al. (2011) used a BG-en iched p oduc (51.4%
BG) om Len inus edodes (insoluble ibe ) as a high- ibe and low-calo ie subs i u e o whea
lou cakes. The esul was inc eased ba e iscosi y wi h mo e shea - hinning beha io and
imp o ed elas ic p ope ies. O e all, he cakes con aining mo e BG showed dec eased olume
and inc eased ha dness, al hough no signi ican di e ences we e obse ed be ween he con ol
and cakes con aining 1 g o yeas β-glucan pe se ing. Heo e al. (2014) also added Len inus
edodes powde ed ex ac (40.1% BG) o imp o e he unc ional p ope ies o glu en- ee ice
noodles. They ound ha he use o such ib ous ma e ial imp o ed he he mo-mechanical
p ope ies o ice lou in a dough sys em, leading o ice noodles wi h g ea e ex ensibili y and
i mness in all h ee concen a ions e alua ed (4, 8, and 12%). Ki isuban e al. (2014) used
esponse su ace me hodology o analyze e ec s o hyd oxyp opyl me hylcellulose (HPMC),
yeas -glucan ex ac (insoluble, 72.2% BG), and whey p o ein isola e (WPI) on physical
p ope ies o glu en- ee b ead baked om o mulas based on ice s a ch. -glucan a ec ed he
speci ic olume di e en ly depending on he WPI le els, i.e., sligh ly inc eased olume wi h -
glucan le els a low WPI le els. This was explained by he abili y o -glucan o inc ease dough
consis ency, which would imp o e gas e en ion and dough de elopmen . Howe e , hey ound
a dec ease in he loa olume o glu en- ee b ead, accompanied by c umb ha dness, a he
highes -glucan le el especially in he p esence o he highes WPI con en . Au ho s ela ed i
wi h oo igid dough ha could cause a limi ed and slow expansion o he gas cells du ing
p oo ing. Tha esul indica ed ha he e was an op imum alue o dough consis ency. The
cons an dough hyd a ion applied by hese au ho s could be, a leas in pa , esponsible o
esul s ob ained. This is because limi ing dough hyd a ion by he addi ion o ing edien s wi h
speci ic wa e abso p ion beha io (such as insoluble-BG-en iched ma e ials) can mask he
impac o i s p esence on dough consis ency, which has well known, d ama ic e ec s on dough
de elopmen abili y (Ronda e al., 2015).
The g ea impac o he solubili y o ibe s on glu en- ee dough heology and b ead making has
been concluded om p e ious s udies (Ma inez e al., 2014). Soluble ibe s (being mainly pu e
ca bohyd a e polyme s such as inulin and polydex ose) dec eased dough consis ency, a o ed
olume inc ease du ing e men a ion, and p oduced b eads wi h highe speci ic olumes, lowe
ha dness, da ke colo , and g ea e cell densi y han con ol b eads. In con as , insoluble ibe s
om oa , bamboo, po a o, and pea, pa icula ly hose o coa se pa icle size, dec eased speci ic
olume and inc eased ma kedly b ead i mness.
The en ichmen o glu en- ee b ead wi h heal h p omo ing ing edien s (which can also
posi i ely a ec he senso ial quali y o his p oduc ) is wo h examining. Consequen ly, he
i s aim o his s udy was o in es iga e he e ec o di e en (1 → 3)(1 → 6)-β-glucan
comme cial concen a es o high pu i y and di e en decla ed wa e solubili y, de i ed om
a ious sou ces, on ice-based dough heology a cons an wa e con en . The sou ces we e
yeas s –soluble powde (SBG) and insoluble powde (IBG)– and ungi – om Pleu o us
Os ea us insoluble powde (FBG). In addi ion, he physical and senso ial p ope ies o BG-
en iched glu en- ee b eads o mula ed a adap ed dough hyd a ion o ge simila consis encies
we e also e alua ed.
2. Ma e ials and me hods
2.1. Ma e ials
Rice lou (12.5% mois u e, 0.46% ash, 7.5% p o ein, 0.49% a , and 79.1% s a ch, pa icle size
dis ibu ion: 6%>150 mm, 150 mm>63.2%>100 mm, and 30.8%<100 mm) was supplied by
He ba Ricemills S.L.U (Ta agona, Spain). Sal , suga , and sun lowe oil we e pu chased om
he local ma ke . The HPMC 4 KM was a gi om Dow Chemical (Midland, Michigan, USA).
(1→3)(1→6)-β-glucans (BG) used in dough and b ead o mula ions we e o h ee ypes. Two o
hem we e ob ained om he yeas Saccha omyces ce e isiae: one wa e insoluble (IBG)
(Wellmune WGP® Dispe sible Powde ) and he o he soluble (SBG) (Wellmune Soluble
Powde ®); Bio he a (Eagan, Minneso a, USA) p o ided bo h as ee samples. Wellmune
WGP® consis s o he pu i ied yeas be a-glucan om he cell wall o bake ’s yeas . The na u al
o m is a small sphe ical pa icle 2-4 mic ons in diame e . The second o m, Wellmune Soluble
Powde ®, has a wide ange o molecula weigh s anging om a ew housand Dal ons o
se e al million Dal ons, wi h an a e age molecula weigh o 100 – 150kDa (in o ma ion
p o ided by he supplie ). The hi d BG, insoluble (FBG), was de i ed om he ungus
Pleu o us os ea us (Pleu an), gi en as a gi by Pleu an, s. .o. (B a isla a, Slo ak Republic);
he a e age molecula weigh o FBG is 760kDa (comme cial in o ma ion). As indica ed by he
supplie s, he p oxima e composi ion o hese p oduc s was: IBG: 79.25% pu i y (d y basis),
84.49% ca bohyd a es, 7.50% a , 4.51% mois u e, 2.86% p o ein, and 0.77% ash; SBG:
91.35% pu i y (d y basis), 92.11% ca bohyd a es, 6.79% mois u e, 0.89% ash, 0.68% p o ein,
and <0.01% a ; FBG: 90.62% pu i y (d y basis), 2.56% mois u e, and absence o lipids and
p o eins.
2.2. Dough p epa a ion
A s aigh dough p ocess was pe o med ollowing he o mula on a 100 g ice lou basis: 92%
wa e , 6% oil, 5% suc ose, 1.8% sal , 2% HPMC, and BG. The le els o pu e BG inco po a ed
in o his o mula ion we e 0 (con ol), 0.5, 1, and 2% in lou basis. Di e en amoun s o
comme cial BG concen a es we e added acco ding o BG pu i y o each ing edien . The
amoun o BG added did no al e he amoun o he o he ing edien s in he o mula ion. The
dough was made by i s blending he solid ing edien s in a ki chen-aid p o essional mixe
(KPM5) o 10 s a speed 2. Then, liquid ing edien s (oil and wa e a 20±2 ºC) we e added and
mixed o 5 min a speed 6 (Pé ez-Qui ce e al., 2014). Each dough was made in duplica e.
2.3. Rheological cha ac e iza ion o dough
2.3.1. Small de o ma ion mechanical es . Oscilla o y and c eep eco e y es s
Oscilla o y and c eep- eco e y es s we e ca ied ou a 25 °C wi h a RheoS ess 1 heome e
(The mo Haake, Ka ls uhe, Ge many) using 60 mm se a ed pa allel pla e geome y. The GF
dough was placed on he heome e pla e using a 3-mm gap and immed, and hen aseline oil
was applied o he ai -exposed su aces o p e en sample d ying du ing es ing. Be o e he
measu emen , he dough was allowed o es o 500 s. A s ess sweep om 0.1 o 200 Pa a 1Hz
was pe o med o es ablish he linea iscoelas ic egion (LVR). F equency sweeps we e ca ied
ou om 10 o 0.1 Hz in he LVR and da a we e i ed o a powe law model as in p e ious
s udies (Ronda e al., 2013). The eco ded iscoelas ic pa ame e s, G1’ and G1’’, and ( an

)1,
ep esen he elas ic and iscous moduli and he loss angen , espec i ely, a a equency o
1Hz. The a, b and c exponen s quan i y he dependence o he dynamic moduli and he loss
angen on he oscilla ion equency. Each es was ca ied ou a leas in duplica e.
C eep es s we e pe o med by imposing a sudden s ep o shea s ess a 1.5 Pa in he LVR o
150 s. In he eco e y phase, he s ess was suddenly emo ed and he sample was allowed o
eco e he elas ic (ins an aneous and e a ded) pa o he de o ma ion o 300 s. Each es was
done in iplica e. Bu ge s model was i ed o c eep and eco e y es s da a, desc ibed in e ms
o compliance J (s ain di ided by he s ess) (Laza idou e al., 2007; Ronda e al., 2013). In his
model, J0 ep esen s he ins an aneous compliance, ela ed o he ins an aneous elas ic dough
de o ma ion when he sample is submi ed o a sudden/cons an s ess; J1 is he e a ded elas ic
o iscoelas ic compliance ela ed o he e a ded dough de o ma ion; and 1 is he e a da ion
ime equi ed o his de o ma ion, ob ained om bo h he c eep and he eco e y phases
(S e e, 1996). η0 is he s eady iscosi y es ima ed om he c eep phase. Jmax is he maximum
c eep compliance ela ed o he maximum dough de o ma ion ob ained a he end o he c eep
s ep. Simila equa ions we e used o he eco e y compliance J ( ). As he e is no iscous low
in he eco e y phase, equa ions consis only o pa ame e s desc ibing he elas ic esponse a e
emo al o he shea s ess. Up o a limi , highe compliances gene ally acili a e highe dough
de elopmen du ing p oo ing and baking (Ronda e al., 2017).
2.3.2. La ge de o ma ion mechanical es : o wa d ex usion es
Fo wa d ex usion assays o o mula ed ice doughs we e done in a TA-XT plus ex u e
analyze (S able Mic o Sys ems, Su ey, UK), equipped wi h a 25-kg load cell and ope a ing a
10 mm/s head speed, ollowing he me hod desc ibed in de ail elsewhe e (Ronda e al., 2015).
Comp ession o ce– ime cu e allowed e alua ing he a e age o ce ob ained om he pla eau,
ep esen ing he o ce necessa y o con inue wi h he ex usion p ocess, and he a ea unde he
cu e, ep esen ing he ene gy needed o he ex usion. Bo h magni udes we e used o de ine
he sample consis ency. All measu emen s we e pe o med in iplica e.
2.4. B ead p epa a ion
B eads we e made in duplica e acco ding he o mula ion desc ibed in Sec ion 2.2. This
included 3 g o d ied yeas pe 100 g o ice lou and a wa e amoun op imized indi idually o

each ype and le el o BG o ob ain he same o ce du ing he ex usion es (see 2.3.2) as wi h
he con ol dough o mula ed wi h 92 g wa e pe 100 g o ice lou . Op imized dough
hyd a ion inc eased wi h ising amoun s o IBG and FBG and dec eased wi h SBG, becoming
be ween 90% and 104% depending on he o mula ion (Table 1). The baking p ocess, desc ibed
in de ail elsewhe e (Pé ez-Qui ce e al., 2014), was ca ied ou in a S eba Dahlen o en (F is ad,
Sweden) a 170 °C o 20 min wi h 7s s eam a he beginning o he baking. A e baking,
b eads we e le o one hou a oom empe a u e be o e analysis. To s udy he e ec on s aling,
b eads we e s o ed a 4±2 °C in polye hylene bags. An expe imen al design o 12 elabo a ions
(see Table 1) was ca ied ou .
Table 1: Amoun s o β-glucan concen a es ob ained om yeas (IBG and SBG) and ungi
(FBG) and dough hyd a ion in each en iched glu en- ee b ead elabo a ion
BG Type
BG (% b*)
WATER (% b*)
Con ol
0
92
SBG
0.5
91.5
SBG
1
91
SBG
2
90
IBG
0.5
95
IBG
1
98
IBG
2
104
FBG
0.5
94
FBG
1
96
FBG
2
100
* b: ice lou basis. SBG: soluble (1-3)(1-6)-β-glucan yeas ; IBG: insoluble (1-3)(1-6)-β-glucan yeas ;
FBG: ungal (1-3)(1-6)-β-glucan (FBG).
2.5. Elec on mic oscope pho omic og aphs o ibe s and c umb b eads
Fibe and b ead c umb pho omic og aphs we e aken wi h a Quan a 200FEI (Hillsbo o, O egon,
USA) en i onmen al scanning elec on mic oscope (ESEM). Fibe pho omic og aphs we e
aken in beam decele a ion mode (BDM) a 2 keV in low acuum mode wi h a backsca e ed
elec on de ec o (BSED). C umb samples we e di ec ly moun ed on s ubs. Obse a ions we e
made wi h an accele a ing ol age o 10 keV.
2.6. E alua ion o b ead quali y
B ead olume was de e mined in ou eplica es by a Volscan p o ile analyze (S able
Mic osys ems, Su ey, UK). B eads we e weighed immedia ely a e emo al om he pan once
cooled. C umb ex u e was de e mined in quad uplica e samples wi h a TA-XT2 ex u e
analyze (S able Mic osys ems, Su ey, UK) using “Tex u e Expe ” so wa e. A 20-mm
diame e aluminum cylind ical p obe was employed in a double comp ession es (TPA) o
pene a e o 50% dep h a 1 mm/s speed es , wi h a 30 s delay be ween i s and second
comp ession. Ha dness (N), chewiness (N), cohesi eness, sp inginess, and esilience we e
calcula ed om he TPA g aph (Gómez e al., 2007). Analyses we e made a 20±3 °C on wo
cen al slices (20 mm hickness) om wo b eads o each dough. B eads we e analyzed esh
and a e one day o s o age a 4 2 °C. Mo eo e , a s aling kine ics s udy was ca ied ou on
b eads en iched wi h he maximum addi ion o BG (2%) by measu ing ex u e a 0, 1, 2, 4, 7,
and 9 days a e baking and s o age a 42 °C. The A ami equa ion was used o i ing he
e olu ion o c umb i mness wi h ime (Ronda and Roos, 2011). C umb and c us colo was
measu ed wi h a Minol a spec opho ome e CN-508i (Minol a Co. LTD., Japan) in he CIE
L*C*h coo dina es using he D65 illuminan , and he 2o s anda d obse e as epo ed elsewhe e
(Ronda e al., 2015). C umb g ain cha ac e is ics o b ead we e assessed by using a digi al
image analysis sys em using ImageJ so wa e. The c umb g ain cha ac e is ics s udied we e he
mean cell a ea (mm2) and he cell densi y (cells/cm2). Loa images we e p e iously acqui ed a
600 do s pe inch wi h a Hp Scanje G3110 scanne (Hewle Packa d En e p ise, Palo Al o, CA,
USA). The analysis was pe o med on 30 x 50 mm squa es aken om he cen e o he loa .
C umb g ain pa ame e s we e measu ed in duplica e.
2.7. Senso y analysis
Senso y analysis was conduc ed on BG-supplemen ed b ead samples using a mul isample
di e ence es ollowing he guidelines sugges ed by Meilgaa d e al. (2007). A ained panel o
eigh panelis s a ed he in ensi y o nine a ibu es on a nume ical in ensi y scale o nine poin s
anging om 1 (no pe cei ed o e y low) o 9 (ex emely in ense o e y high). The con ol
sample was used as a e e ence and was posi ioned in he middle o he scale (Ronda e al.,
2005). The samples we e e alua ed in e ms o c us uni o mi y, c umb g ain uni o mi y, odo
and la o in ensi y, a e as e pe sis ency, c umb humidi y, c umb adhesi eness, c umb
so ness, and c umb cohesi eness. Each a ibu e was p esen ed sepa a ely.
2.8. S a is ical analyses
S a g aphics Cen u ion .16 (Bi s eam, Camb idge, MN, USA) was used o non-linea
eg essions o i c eep- eco e y da a o Bu ge s model. ANOVA analysis, LSD (Leas
Signi ican Di e ence) es (p<0.05), and Pea son co ela ion analysis we e pe o med using he
.6. S a is ica package (Tulsa, OK, USA). The wo ac o s s udied in ANOVA analysis we e he
ype o (1 → 3)(1 → 6)-β-glucan added o he dough and i s addi ion le el. In he senso y
analysis, he e ec o panelis was also checked.
3. Resul s and discussion
3.1. BG ing edien mic os uc u e
Fig. 1 shows he pho omic og aphs o he di e en BG-con aining ing edien s used in his
s udy. BG om yeas exhibi ed sphe ical o sligh ly len icula mo phologies esponding o i s
o igin om yeas cells, while SBG showed a smoo h su ace, and IBG, a g ainy su ace. The
pa icles o FBG e lec ed he c ushed ma ix o mush oom issue, showing an i egula su ace
and polyhed al shape, and seemed o be mo e disin eg able. All pa icles o BG ing edien s
showed an a e age pa icle size below 50 m, which means ing edien s o ine s uc u e.
3.2. Viscoelas ic p ope ies o ibe -en iched doughs a cons an dough hyd a ion
Dough p epa ed wi h ei he cons an consis ency o cons an wa e addi ion has been used o
es ing he e ec o added subs ances. The la e , p e iously used by K upa-Kozak e al. (2012)
and Nunes, Ryan, and A end (2009), was selec ed in his s udy o dough heology
cha ac e iza ion because eco ded di e ences in dough beha io may be di ec ly ela ed o he
added ing edien s. Mo eo e , his app oach allows be e objec i i y in he compa ison o he
a ailabili y o wa e o s a ch gela iniza ion.
The iscoelas ici y o glu en- ee doughs was examined by oscilla o y and c eep measu emen s.
Table 2 shows he pa ame e s ob ained om i ing equency sweeps and c eep es da a o
Powe law and Bu ge s model, espec i ely. The R2 alues o he i ing o powe law o
equency sweep esul s we e always abo e 0.99. The i ing o Bu ge s model o c eep-
eco e y es da a led always o R2 alues abo e 0.97. All heological p ope ies o doughs we e
signi ican ly (p<0.05) a ec ed by he ype o he ing edien used and he le el o addi ion. The
double in e ac ion ( ype o BG*le el) also had a signi ican e ec on all heological dough
p ope ies, excep o he exponen s a, b and he phase shi angen ; his indica es ha he le el
o addi ion had a di e en e ec depending on ing edien ype in mos iscoelas ic p ope ies. In
all cases he s o age modulus (G’) was much la ge han he loss modulus (G’’), implying he
p e alence o elas ic ea u es o e iscous, sugges ing a ypical weak gel s uc u e, in ag eemen
wi h p e ious s udies on GF dough en ichmen wi h ce eal BG (Hage e al, 2011; Laza idou e
al., 2007; Ronda e al., 2013, 2015). Bo h moduli showed sligh inc eases wi h angula
equency as e idenced by he low a and b exponen s. The elas ic and iscous moduli a 1 Hz,
G1’, and G1’’ dec eased as soluble p oduc SBG was added, o 30% a he highes addi ion le el,
deno ing a so e dough consis ency. This has been obse ed by Ziob o e al. (2013) and
Pe essini e al. (2015) wi h he addi ion o inulin o glu en- ee and whea doughs, espec i ely.
Figu e 1. Scanning elec on mic oscope pho omic og aphs o he comme cial (1→3)(1→6)-β-glucan p oduc s: (a) FBG. (b) IBG. (c) SBG; and o he glu en-
ee b eads: (d) Con ol b ead (wi hou BG). (e) B ead wi h 2% FBG. ( ) B ead wi h 2% IBG. (g) B ead wi h 2% SBG. The inse s included in a), b) and c)
show a de ail o he BG pa icula e su ace.
a
b
c
d
e
g
s uc u e. An imp o ed c umb was p e iously obse ed wi h he addi ion o oa BG o bo h GF
b eads (Laza idou e al., 2007; Ronda e al., 2015) and whea b eads (Wang, Mille , and
Hoseney, 1998). This e ec o oa BG addi ion was ela ed o he capaci y o his soluble ibe
o p e en coalescence o he cells du ing dough p oo ing and baking.
The BG-p oduc -en iched b eads showed c us s wi h signi ican ly highe L* and C* coo dina es
han hose o he con ol sample. This means ha BG- o i ied b eads we e ligh e and mo e
i idly colo ed han he con ol b ead. The highe dough wa e con en (in he case o adding an
insoluble ing edien ) and he addi ion o a wa e - e aining ing edien , such as BG, dec eased he
Mailla d eac ions a e and yielded a ligh e c us (Pe ez-Qui ce e al., 2014). This has
p e iously been no ed by Hage e al. (2011) and Ronda e al. (2015) o ce eal BG. BG
mode a ely a ec ed he c us hue, h, depending on BG solubili y: IBG dec eased i , p oducing a
eddish c us ; while SBG inc eased i , leading o yellowish ones. En iched b eads showed
sligh ly da ke c umbs (lowe L*). Likewise, in he case o insoluble ing edien s, en iched
b eads showed signi ican ly lowe Ch oma (IBG) and hue (FBG), deno ing less i id and
eddish c umbs, espec i ely. The sligh e ec o ibe on b ead c umb colo could be ela ed o
he o iginal –al hough e y sligh – colo o hese ing edien s.
Figu e 3 p esen s c umb i mness e olu ion du ing he s o age o BG-added GF b eads a 42
°C. The co ela ion coe icien s o A ami model i ing o expe imen al da a, R2, anged om
0.96 ( o FBG) o >0.99 ( o he es ). The alues o he A ami model pa ame e s (Fo, F, k, n)
we e (0.59N, 2.5N, 0.59 d-n, 0.37), (0.48N, 2.2 N, 0.93 d-n, 0.19), (0.38N, 2.3 N, 0.35 d-n, 0.50),
and (0.80N, 3.4 N, 0.39 d-n, 0.69) o SBG-, IBG-, FBG-added b eads, and con ol b ead,
espec i ely. The hal -li e ime, 1/2, epo s he ime equi ed o achie e 50% o le eling-o
ex en o he i mness. The hal -li e ime was 1.5, 0.2, 3.9, and 2.3 days, espec i ely, meaning
as e ha dening du ing s o age o yeas BGs-added c umbs. The a e cons an s o c umb
i ming ound in his s udy we e o he same o de o magni ude as ha ound in o he s udies
o glu en- ee b eads, 0.44 d-n (Ronda and Roos, 2011), and inulin-en iched whea b eads, 0.35
d-n (Ronda e al., 2014), s o ed a 42 °C. F om he i s days, FBG-added b eads showed he
lowes c umb i mness, while b eads wi h yeas BG ibe (IBG and SBG) showed i mness a e
one day o s o age simila o ha o he con ol b ead. Howe e , he lowe le elling-o i mness
alues, F, o ibe -added b eads ( anging om 2.2 o 2.5N e sus 3.5N o he con ol b ead)
and he esul s p esen ed in Fig. 3 show ha BG ibe (especially FBG) helped educe b ead
ha dening a e long- e m s o ages. The pa ame e Fo was simila o alues o i mness
measu ed in esh b eads (Table 3), which con i ms he good i o he A ami model o
expe imen al da a. The highe a e cons an , k, and he lowes hal -li e ime, 1/2, ob ained o

IBG-added b eads indica e a as e change in i mness ( om he ini ial esh b ead alue o he
le elling-o one) in compa ison wi h he emaining b eads.
Figu e 3: Ha dening kine ics in BG-en iched b eads s o ed a 4  2 ºC. The con inuous lines
esul ed om i ing he A ami equa ion o expe imen al da a. The e o ba s ep esen he
s anda d de ia ion.  Con ol;  SBG  IBG  FBG
3.5. Senso y e alua ion
The panelis e ec on senso y e alua ion was p e iously e i ied and was no signi ican
(p>0.05) o any senso y a ibu es. Panelis s we e unable o ind signi ican (p<0.05)
di e ences be ween BG-en iched and con ol samples in e ms o c umb humidi y, c umb
adhesi eness, c umb so ness and c umb cohesi eness (da a no shown). Simila esul s we e
ob ained p e iously by Ma ins e al. (2015), who s udied BG-b ead o i ica ion wi h d y spen
yeas ; hey epo ed no signi ican di e ences in senso ial a ibu es o he inal p oduc . An
analogous beha io was also ound in ibe -en iched p oduc s such as inulin- o i ied snacks
(Pe essini e al., 2015). Figu e 2 shows he senso y e alua ion esul s o BG-supplemen ed
glu en- ee b eads in e ms o c us uni o mi y, c umb g ain uni o mi y, odo and la o
in ensi y, and a e as e pe sis ency, in compa ison o he con ol b ead (which was posi ioned in
he middle o he scale). SBG-supplemen ed b ead a a le el o 0.5-2% showed a senso ial
p o ile e y simila o con ol b ead excep o c us uni o mi y. This pa ame e signi ican ly
(p<0.05) inc eased when SBG was added, p obably due o he highe olumes ob ained in he
o i ied b eads. Con e sely, adding IBG and FBG esul ed in lowe sco es han hose o he
con ol sample, deno ing ha he loa es looked wo se and he c umb g ain was less uni o m.
Insoluble-BG- o i ica ion led o highe la o and odo in ensi y (mainly o IBG) and
a e as e ( o bo h IBG and FBG) sco es han con ol b ead. Panelis s indica ed ha hese
di e ences s emmed om a sligh s ange, undesi able as e and smell.
4. Conclusions
This s udy demons a ed he easibili y o en iching glu en- ee b ead wi h (1 → 3)(1 → 6)-β-
glucan en iched p oduc s ob ained om yeas o ungi a 0.5-2% le el. The e ec on dough
heological p ope ies was e y dependen on BG sou ce and solubili y. In gene al, he soluble
en iched p oduc es ed dec eased G’, G’’, and s eady iscosi y and inc eased compliances.
Howe e , doughs wi h insoluble ibe s gene ally showed he opposi e end, inc easing hei
esis ance o de o ma ion as he concen a ion inc eased. The physical quali y o b eads was
signi ican ly imp o ed by adding (1→3)(1→6)-β-glucan a op imized dough hyd a ion. The
addi ion o (1→3)(1→6)-β-glucan p oduc s caused an inc ease in he speci ic olume o he
b eads and educed hei ha dness, while leading o delayed c umb ha dening du ing s o age.
Senso y e alua ion also demons a ed an imp o emen in b ead o ganolep ic a ibu es when
soluble BG was added. Insoluble BG ex ac s, bo h om yeas o ungi, a 1% and 2% le els,
ga e b eads a e y sligh s ange, undesi able as e and smell no ound in he con ol ice lou
b ead. The use o a mo e complex, op imized o mula ion could help o e come his sligh ly
undesi able as e/odo . Addi ional s udies a e s ill pending in his sense.
Acknowledgemen s: The au ho s g a e ully acknowledge he inancial suppo o he Spanish
Ins i u ion Minis e io de Economía y Compe i i idad and he Eu opean Regional De elopmen
Fund (FEDER) (P ojec s AGL2012-35088 and AGL2015-63849-C2-2-R) and Conseje ía de
Educación (Jun a de Cas illa y León) / FEDER (P ojec VA072P17). Au ho s hank D . Joanna
Ha asym o he e ision o he pape and he commen s abou ungi and yeas (1→3)(1→6)-β-
glucans ex ac ion and p ope ies. Ma ina Villanue a hanks he Jun a de Cas illa y León o he
doc o al g an .
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