Rheological and microstructural evolution of the most common gluten-free flours and starches during bread fermentation and baking

Accep ed Manusc ip Rheological and mic os uc u al e olu ion o he mos common glu en- ee lou s and s a ches du ing b ead e men a ion and baking Ma io M. Ma ínez, Manuel Gómez PII: S0260-8774(16)30407-1 DOI: 10.1016/j.j oodeng.2016.11.008 Re e ence: JFOE 8713 To appea in: Jou nal o Food Enginee ing Recei ed Da e: 24 June 2016 Re ised Da e: 11 Sep embe 2016 Accep ed Da e: 9 No embe 2016 Please ci e his a icle as: Ma ínez, M.M., Gómez, M., Rheological and mic os uc u al e olu ion o he mos common glu en- ee lou s and s a ches du ing b ead e men a ion and baking, Jou nal o Food Enginee ing (2016), doi: 10.1016/j.j oodeng.2016.11.008. This is a PDF ile o an unedi ed manusc ip ha has been accep ed o publica ion. As a se ice o ou cus ome s we a e p o iding his ea ly e sion o he manusc ip . The manusc ip will unde go copyedi ing, ypese ing, and e iew o he esul ing p oo be o e i is published in i s inal o m. Please no e ha du ing he p oduc ion p ocess e o s may be disco e ed which could a ec he con en , and all legal disclaime s ha apply o he jou nal pe ain. MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 1 Rheological and mic os uc u al e olu ion o he mos common glu en- ee lou s 1 and s a ches du ing b ead e men a ion and baking 2 3 Ma io M. Ma ínez 1,2,* , Manuel Gómez 1 4 5 1 Food Technology A ea. College o Ag icul u al Enginee ing, Uni e si y o Valladolid, 6 Palencia, Spain 7 2 P esen add ess: Whis le Cen e o Ca bohyd a e Resea ch, Depa men o Food 8 Science, Pu due Uni e si y, Wes La aye e, IN 47907, U.S.A. 9 10 * Co esponding au ho e-mail: ma[email p o ec ed]om 11 ma 1269@pu due.edu 12 13 14 15 16 17 18 19 20 21 22 23 24 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 2 Abs ac 25 Mechanis ic ela ions be ween he e olu ion o he s a ch/ lou s uc u e, dough 26 heology and b ead quali y we e in es iga ed using he mos common lou s and 27 s a ches in glu en- ee b ead-making. Mic og aphs showed ha he small whea s a ch 28 g anules illed he spaces o he big g anules, o ming a uni o m s a ch-hyd ocolloid 29 ma ix. This g anula ad an age dec eased he consis ency and inc eased he uni o mi y 30 o whea -s a ch based doughs h oughou e men a ion, as shown by mic og aphs and 31 he highe c i ical s ain. The iscoelas ic p ope ies o he di e en doughs s ongly 32 in luenced he b ead olume and he c umb ex u e. Thus, s a ch-based b eads showed 33 highe speci ic olume and lowe ha dness, especially hose made wi h whea s a ch, 34 whose lowe pas ing empe a u e also ein o ced he con inuous phase o he c umb. On 35 he o he hand, he la ge po a o s a ch g anules did no o m a con inuous s a ch-36 hyd ocolloid ma ix, esul ing in b eads wi h he lowes speci ic olume, elas ici y, 37 cohesi eness and esilience, and he highes ha dness. 38 39 Keywo ds: s a ch, lou , glu en- ee, b ead, heology, mic os uc u e 40 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 3 1. In oduc ion 41 Glu en plays a p incipal ole in b ead de elopmen by gi ing cohesi eness and 42 p omo ing he e en ion o he CO 2 p oduced du ing e men a ion. Thus, gas expansion 43 causes whea b eads o gain olume and a ain accep able c umb ex u e (Deo a e al., 44 2014). Recen ly, he ma ke o glu en- ee b eads has expanded and subs an ial e o s 45 a e unde way o enhance hei quali y. 46 In whea -con aining doughs, heological s udies a e c ucial o unde s anding he 47 unc ionali y o lou s and addi i es as well as p edic ing he dough machinabili y and 48 b ead quali y (S ojceska and Bu le , 2012). The small ampli ude oscilla o y shea 49 (SAOS) echnique is ideal o cha ac e ize he s uc u al p ope ies o iscoelas ic 50 ma e ials (Mo ison, 2001). In he las decade, c eep- eco e y has become ano he 51 echnique used o cha ac e ise he s uc u al p ope ies o iscoelas ic doughs. I 52 comp ises a s a ic heological me hod in which an ins an aneous s ess is applied o he 53 sample and he change in s ain is measu ed o e ime. A c eep phase is usually 54 ollowed by a eco e y phase in which he applied s ess is emo ed (S e e, 1996). 55 S udies connec ing glu en- ee dough heology wi h he quali y o he esul an b ead 56 a e sca ce. While i is ue ha nume ous wo ks include heological analyses o dough 57 cha ac e iza ion, mos ly comp ising SAOS and in lesse ex en la ge de o ma ion 58 analyses (Masu e e al., 2016), he e a e s ill no uni e sal indica o s ha well co ela e 59 glu en- ee dough heology wi h he quali y o he esul an b ead. In some wo ks, an 60 inc ease in he b ead olume was app ecia ed as iscoelas ic moduli (G’ and G’’) 61 dec eased (Mancebo e al., 2015a,b; Rocha-Pa a e al., 2015). Howe e , s udying 62 di e en hyd ocolloids, Mancebo e al. (2015b) obse ed ha he c eep- eco e y 63 echnique could be mo e sui able han oscilla o y shea es s o p edic b ead olume. 64 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 4 The absence o a eliable heological indica o can be a ibu ed o he di e se 65 heological e olu ion o he di e en doughs du ing p ocessing ( e men a ion and 66 baking). Rheological a ibu es o whea doughs s a changing a he beginning o 67 e men a ion. This is mainly due o: 1) he CO 2 expansion p e iously o med wi hin he 68 gas cells and, 2) he pH modi ica ion by means o such CO 2 and i s in luence on he 69 glu en ne wo k (Pyle and Go on, 2008). In glu en- ee b ead making, wi hou a glu en 70 ne wo k sensi i e o acidi ica ion, non-s udied simila heological phenomena could be 71 p oduced. As o he baking s ep, he e men ed dough is exposed o hea ans e om 72 he o en, esul ing in a chain o phenomena go e ned by hea and mois u e ans e s 73 (Le-Bail e al., 2011). A i s he hea ans e esul s in an expansion o he gas cells 74 con ained in he e men ed dough ia: 1) inc eased CO 2 p oduc ion by yeas (un il yeas 75 inac i a ion a 50-60°C), 2) gas expansion, 3) apo iza ion o he CO 2 and solubilized 76 e hanol in he liquid phase o he dough and 4) mois u e apo iza ion (Zhang, Lucas, 77 Dou sa , Flick and Wagne , 2007). Howe e , when eaching a ce ain empe a u e, he 78 hyd a ed s a ch gela inizes and he p o ein coagula es, leading o c umb se ing (Le-Bail 79 e al., 2011), and he e o e o an amo phous s uc u e ha co e s he gas cells. In 80 pa icula , whea , co n, ice and po a o s a ches ha e been epo ed o ha e 81 gela iniza ion empe a u e anges o 58-64, 62-72, 68-78 and 58-68°C, espec i ely 82 (Biliade is 2009). This amo phous ma ix, o med mainly by gela inized s a ch, will be 83 u he modi ied du ing cooling as s a ch e og ades, in luencing b ead ex u e. 84 The mos commonly used s a ches in glu en- ee b ead-making a e maize s a ch and 85 some s a ches om ube s, such as po a o and apioca (Masu e e al., 2016), despi e he 86 g owing p ominence o he gua an eed glu en- ee whea s a ch in he las ew yea s 87 (Mancebo e al., 2015a). As o he lou s, ice lou is he mos commonly used, 88 ollowed by maize lou , since hey a e he mos highly p oduced and a o dable ce eals. 89 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 5 To he bes o ou knowledge, mechanis ic s udies, showing he in luence o mos 90 commonly used s a ch-based ing edien s in glu en- ee b ead-making on he in e play 91 be ween dough heology and b ead quali y, a e sca ce. 92 S a ches and lou s ha e ex ensi e mic os uc u al di e ences a g anula s uc u al 93 scales, which can in luence hei capaci y o gene a e glu en- ee b eads wi h high 94 quali y s anda ds. Howe e , use o scanning elec on mic oscopy (SEM) as a ool o 95 iew glu en- ee doughs and b eads ha e been epo ed on e y ew occasions (de la 96 He a e al., 2013; Ma inez e al., 2014; O´Shea e al., 2013; Pe essini e al., 2011; 97 Yano, 2010). In gene al, hese s udies a e based on he use o a single glu en- ee 98 lou /s a ch o hei combina ion, i.e., al oge he du ing he mixing p ocess. 99 Ne e heless, compa a i e s udies on he single e ec o di e en s a ches and lou s 100 a e sca ce, and none o hem include heological and mic os uc u al analysis. 101 The objec i e o his s udy was o ob ain a compa a i e insigh o he e olu ion o he 102 mos common lou s and s a ches used in glu en- ee b ead making du ing e men a ion 103 and baking. In his way, changes p oduced in he doughs a la ge s uc u al scales we e 104 pic u ed h ough SEM du ing e men a ion and ela ed o he e olu ion o he dough 105 iscoelas ici y (SAOS and c eep- eco e y). In addi ion, he de elopmen o he b ead 106 olume and c umb ex u e and mic os uc u e du ing baking was also s udied. We 107 belie ed ha esul s could show mechanis ic co ela ions be ween he de elopmen o 108 he s a ch/ lou s uc u e, dough heology and b ead quali y, gi ing aluable 109 in o ma ion wi h he aim o p edic ing he quali y o he esul an glu en- ee b ead. 110 111 2. Ma e ials and me hods 112 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 6 2.1. Ma e ials 113 Coa se ice lou and maize lou we e supplied by Ha ine a Cas ellana SL (Medina del 114 Campo, Spain) and Maice as Españolas, S.A. (Valencia, Spain), espec i ely. Whea 115 and po a o s a ch we e p o ided by Roque e (Les em, F ance) whe eas Miwon maize 116 s a ch (Daesang Co., Seoul, Ko ea) was pu chase om he local ma ke . The es o 117 ing edien s used o b ead-making we e VIVAPUR 4KM HPMC (Hyd oxyp opyl 118 Me hylcellulose, JRS, Rosenbe g, Ge many), Sa -Ins an d y yeas (Lesa e, Lille, 119 F ance), sal (Unión Saline a de España, Mad id, Spain), suc ose (Azuca e a, AB, 120 Mad id, España), sun lowe ABRISOL (Ou ense, Spain) and ap wa e . 121 Flou and s a ch composi ion was de e mined using he AACC me hods (AACC, 2015) 122 44-15.02 (mois u e con en ) and 46-30.01 (p o ein) wi h a Leco T uSpec de ice (Leco, 123 S . Joseph, MI, USA). The mos ou s anding physical p ope ies o he di e en lou s 124 and s a ches we e also cha ac e ised o be e unde s and he heological and 125 mic os uc u al beha iou du ing e men a ion and baking. Pa icle size was measu ed 126 wi h a lase di ac ion pa icle size analyse (Mas e size 3000, Mal e n Ins umen s, 127 L d., Wo ces e shi e, UK). The mean diame e o equi alen olume o mass d(4,3), 128 which indica es he cen al poin o he olume dis ibu ion o he pa icles, was 129 eco ded. Wa e binding capaci y, de ined as he amoun o wa e e ained by he 130 lou /s a ch a e being subjec ed o cen i uga ion, was measu ed as desc ibed in he 131 me hod 56-30.01 (AACC 2015). The pas ing p ope ies we e analysed using he 132 s anda d me hod 61-02.01 (AACC, 2015) wi h a Rapid Visco Analyse (RVA-4) 133 (Pe en Ins umen s Aus alia, Macqua ie Pa k, Aus alia). These analyses we e ca ied 134 ou in duplica e. Da a a e shown in Table 1. 135 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 7 2.2. Me hods 136 2.2.1. Dough p epa a ion and b ead-making 137 The ollowing ing edien s we e used in b ead-making: wa e (100 g/100 g lou o 138 s a ch), ins an d y yeas (3 g/100 g), sal (1.8 g/100 g), oil (6 g/100 g), HPMC (2 g/100 139 g) and whi e suga (5 g/100 g). In all es s, he wa e empe a u e was held be ween 20 140 and 22 °C. Yeas was p e iously dissol ed in he wa e be o e i s inco po a ion. All he 141 ing edien s we e mixed o 8 min in a Ki chen Aid 5KSM150 mixe (Ki chen Aid, 142 Michigan, USA) wi h a dough hook (K45DH) a speed 2. Fe men a ion was pe o med 143 a 30 °C and 80 % RH o 90 min. A e e men a ion, doughs we e baked in an elec ic 144 modula o en o 40 min a 190 °C. B ead-making was pe o med in duplica e. 145 Fo dough e alua ion, 100 g o dough ob ained a e mixing, 45 min and 90 min o 146 e men a ion we e placed in small aluminium moulds (140x40x35 cm, ALU-Schale, 147 Wikla n, Ge many), in oduced in o polye hylene plas ic bags and immedia ely ozen 148 a -21°C. Doughs we e kep in he eeze du ing 24 hou s be o e heological and 149 mic os uc u al analyses. 150 Fo b ead cha ac e iza ion, 250g o dough ob ained a e mixing we e placed in 151 aluminium moulds (232x108x43.5 cm, ALU-Schale, Wikla n, Ge many) and hen 152 e men ed and baked ollowing he baking desc ibed abo e. B eadswe e aken ou om 153 he o en a e 20 and 40 min o baking. Subsequen ly, he loa es we e emo ed om 154 he moulds a e a 60-min cooling pe iod. They we e hen in oduced in o polye hylene 155 plas ic bags and s o ed a -21 °C du ing 24h un il analysis. 156 2.2.2. Mic os uc u al analysis o doughs and b eads 157 Dough and b ead pho omic og aphs we e aken wi h Quan a 200FEI (Hillsbo o, 158 O egon, USA) en i onmen al scanning elec on mic oscope (ESEM). Pho omic og aphs 159 we e aken in high acuum mode. C umbs pic u es we e aken om a pe pendicula 160 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 8 slan o he cell wall, i.e., showing he su ace o a gas cell wall. Con e sely, c us 161 pic u es we e aken showing hei leng hwise sec ion, in o he wo ds, highligh ing he 162 hickness o he c us . 163 2.2.3. Rheological p ope ies o doughs 164 Be o e conduc ing any heological measu emen , doughs we e allowed o es in he 165 measu emen posi ion o 10 min as equilib a ion ime, i.e., he necessa y ime o allow 166 he s esses induced du ing sample loading o elax. The equi ed equilib a ion ime was 167 selec ed acco ding o p e ious ime sweep es s ca ied ou wi hin he linea egion (1 168 Pa) a 1 Hz and 25°C du ing 30 min. The ime sweep es showed ha in less han 10 169 min alues o G’ and G’’ became independen o ime. A e adjus men o he gap, he 170 excess dough was emo ed and he exposed edges o he samples we e always co e ed 171 wi h aseline oil (Pan eac Química S.A., Cas ella del Valles, Spain) o a oid sample 172 d ying du ing measu emen s. In his s udy, yeas -con aining doughs we e analyzed a e 173 kneading (0min o e men a ion), 45 and a 90 min o e men a ion in o de o include 174 he e ec s o he gas olume and e men a ion me aboli es. All heological es s we e 175 un in duplica e in a con olled s ess a heome e (Haake RheoS ess 1, The mo Fische 176 Scien i ic, Sche e e, Ge many) wi h a i anium pa allel pla e geome y senso PP60 Ti 177 (60 mm diame e , and 3 mm gap). 178 2.2.3.1. Viscoelas ic p ope ies 179 Linea iscoelas ic p ope ies we e s udied by small ampli ude oscilla o y es (SAOS). 180 Dynamic linea iscoelas ic ange was es ima ed by pe o ming a s ess sweep om 0.1 181 o 50 Pa a a equency o 1 Hz. 182 F equency dependence expe imen s we e conduc ed om 10 o 0.01 Hz a 25 °C. The 183 applied s ess was always selec ed o gua an ee he exis ence o linea iscoelas ic 184 esponse. A leas wo eplica es o each oscilla o y shea es we e conduc ed. 185 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 15 which is in ag eemen wi h he obse ed shi o he c osso e o lowe equencies. As 335 men ioned, his can sugges a g adual inc ease o he CO 2 bubble packing and a lowe 336 in ensi y o HPMC en anglemen s (see also Fig. 1). 337 3.1.4. C eep- eco e y es 338 The abili y o doughs o eco e some s uc u e by s o ing ene gy was analysed by 339 applying an ins an aneous s ess and measu ing he change in s ain o e ime (Fig. 2). 340 This was pe o med as a seconda y analysis o he dough elas ici y. The c eep eco e y 341 cu es o glu en- ee doughs exhibi ed a ypical iscoelas ic beha iou combining bo h 342 iscous luid and elas ic esponses (Laza idou e al., 2007). Doughs made wi h lou s 343 exhibi ed lowe compliance alues in bo h c eep and eco e y phases. This occu ence 344 is in ag eemen o wha was obse ed in ano he s udy compa ing ice lou wi h o he 345 s a ches (Mancebo e al., 2015a) and in he mechanical spec a o he cu en wo k. 346 Again, his would indica e highe dough consis ency (Edwa ds e al., 2003). Among 347 s a ches, whea s a ch displayed highe compliance alues han maize s a ch a he h ee 348 e men a ion imes, which is in ag eemen wi h he low consis ency (low iscoelas ic 349 moduli) o whea s a ch doughs obse ed in he mechanical spec a. As o he po a o, a 350 di e en end was exhibi ed, wi h he highes compliance alues a ime 0 o 351 e men a ion. Howe e , in his case, and con e se o he es o he samples, a s ong 352 inc ease o he compliance as he e men a ion p oceeded was no obse ed. This e en 353 could be due o he la ge size o po a o g anules wi h he absence o supe icial po es, 354 which could make dough less e icien in e ms o g anule packing and o ming a 355 con inuous phase. 356 An addi ional pa ame e ha can be ex ac ed om he c eep eco e y es is he 357 di e ence be ween he compliance alue a he e minal egion o he cu e, whe e 358 dough eco e y has eached equilib ium, and he maximum compliance eached a he 359 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 16 end o he c eep phase, called s eady s a e compliance (J e ) (Laza idou e al., 2007). This 360 alue is an indica o o he elas ici y o he dough. In Fig. 2, highe s eady s a e 361 compliance is obse ed o doughs made wi h whea s a ch, which could be explained 362 h ough he mechanisms discussed in he p e ious sec ions. 363 3.2. Physical and mic os uc u al e olu ion o b eads du ing baking 364 3.2.1. Mic os uc u al e olu ion o b ead c umb 365 Du ing baking, he s uc u al and physical p ope ies o b ead change, whe ein 366 semisolid dough ans o ms o b ead wi h so inne c umb and c ispy ou e c us . The 367 magni ude o hese ans o ma ions in glu en- ee b eads will especially depend on he 368 s a ch p ope ies. The c umb de elopmen o he di e en b eads du ing baking was 369 isually moni o ed h ough SEM (Fig. 4). In all he samples, images we e aken 370 pe pendicula ly o he cell walls o obse e hei su ace. All pic u es showed he 371 p esence o a con inuous ma ix o med by he s a ch and hyd ocolloid, bu in con as 372 o dough mic og aphs, he g anules we e mo e igh ly compac ed. Nume ous 373 physiochemical and biological ans o ma ions, mainly CO 2 elease, gas olume 374 expansion, wa e e apo a ion and s a ch gela iniza ion, ake place du ing b ead-baking 375 p ocess (Chhanwal and Anandha amak ishnan, 2015). Doughs made wi h s a ches 376 p esen ed a mo e uni o m con inuous phase han lou -based c umbs, especially hose 377 made wi h whea s a ch. I seems ha he building and packing ea u es o he bimodal 378 sized whea s a ch oge he wi h i s lowe pas ing empe a u e (Figs. 1, 2, 3) con ibu ed 379 o c ea e a con inuous phase ha , a e gela iniza ion, will lead o a con inuous c umb 380 s uc u e (p ecu so o an accep able c umb cohesi eness and esiliency). I is 381 no ewo hy ha he la ge s a ch g anules obse ed in he po a o sample s ill looked 382 pe ec ly ounded, indica ing ha hey p obably we e no ully gela inised du ing 383 baking. As he cou se o baking p og essed, he empe a u e inc ease ini ia ed wa e 384 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 17 e apo a ion and ca bon dioxide elease, which esul ed in o en sp ing du ing ini ial 385 baking s age. Ca bon dioxide elease igge ed he uppe expansion o he op c us and 386 concu en ly he de elopmen o c umb. S uc u al changes occu du ing he whole 387 b ead-baking p ocess and hey comp ise mainly solidi ica ion and expansion. The 388 ne wo k-like s uc u e o b ead c umb is p edominan ly due o s a ch gela iniza ion 389 (Zhou and The d hai, 2007), as shown in Fig. 4. 390 The de elopmen o he c us mic os uc u e du ing baking was also s udied. The 391 doughs made wi h lou s exhibi ed a s uc u e o med by he s a ch g anules su ounded 392 by a p o ein ma ix in which in ac lou pa icles we e s ill isible. On he o he hand, 393 he c us sec ion o doughs made wi h s a ch appea ed sligh ly less uni o m. 394 Mic og aphs also showed ha s a ch did no gela inize, o ming a compac ex e nal 395 laye . In he c us , wa e e apo a es quickly, lea ing he s a ch wi h no a ailable wa e 396 o gela iniza ion. In addi ion, s eam was no applied a he beginning o baking, which 397 was al eady epo ed o p omo e s a ch gela iniza ion in he c us (Al ami ano-Fo oul e 398 al., 2012; Le-Bail, e al., 2011). Howe e , signi ican changes we e no isible and a 399 clea end was no obse ed (Supplemen a y ma e ial). 400 3.2.2. Physical p ope ies o b eads 401 The e ec o he ype o s a ch sou ce and he baking ime on he speci ic olume and 402 c umb ex u e is shown in Table 3. B eads made wi h lou s had less speci ic olume 403 han hose made wi h s a ch. This could be ela ed o he high consis ency o lou -based 404 doughs, i.e., high iscoelas ic moduli and low maximum c eep compliance (Ma inez e 405 al., 2015b). As men ioned, i can be a ibu ed o he bigge pa icle size and he 406 p esence o a p o ein laye obse ed in Fig. 1. In pa icula , b eads made wi h maize 407 lou exhibi ed he lowes speci ic olume, which could be due o he highe wa e 408 abso p ion capaci y o maize lou compa ed o ice lou (Table 1). Meanwhile, s a ch-409 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 18 based b eads showed a highe speci ic olume, especially whea s a ch-based b eads, 410 ollowed by he b ead made wi h maize s a ch. This is in ag eemen wi h he p e ious 411 esul s ob ained in he heological analysis, whe e whea s a ch-based doughs had lowe 412 consis ency, i.e., lowe iscoelas ic moduli (Fig. 2), be e packing p ope ies and 413 capaci y o o m a uni o m con inuous ma ix in he dough (Figs. 1, 4). In addi ion, he 414 lowe pas ing empe a u e indica es ha whea s a ch s a s o gela inize ea lie , 415 leaching amylose ha could inc ease he iscosi y and elas ici y o he con inuous 416 s a ch-hyd ocolloid con inuous phase (Table 1). Also in good co ela ion wi h he 417 heological and mic os uc u al analysis, po a o s a ch-based b eads had he lowes 418 speci ic olume among he s a ch-based b eads. This occu ence can be a ibu ed o he 419 la ge g anula size o po a o s a ch, which p e en s he s a ch om o ming an 420 accep able con inuous phase wi h he es o he dough/c umb componen s. 421 Speci ic olume was in e sely co ela ed wi h c umb ha dness. This ecip ocal 422 ela ionship has been epo ed in p e ious s udies on glu en- ee b ead (Gallaghe e al., 423 2003), and i was a ibu ed o he lowe esis ance o dough de o ma ion, wi h a highe 424 pe cen age o ai con en . In gene al, s a ches showed a so e c umb wi h highe 425 elas ici y and esilience han lou s. Again, whea s a ch c umbs showed he bes 426 ex u al p ope ies (lowe ha dness and highe elas ici y, cohesi eness and esilience), 427 likely a ibu ed o he con ibu ion o he whea s a ch s uc u e.. 428 The de elopmen o he olume and ex u al pa ame e s o b eads along he cou se o 429 e men a ion is also shown (Table 3). C umb elas ici y, cohesi eness and esilience 430 we e no changed om 20 min o he end o e men a ion. Acco ding o he esul s, i 431 seems ha some a ibu es o c umb s uc u e a e o med a he ea ly s age o he 432 e men a ion and hen hey emain cons an . Howe e , b ead olume inc eased o e 433 e men a ion, leading o so e c umbs, indica ing ha some changes occu du ing he 434 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT 19 en i e baking p ocess. These changes p oduced in he s uc u e o b ead c umb a e 435 p edominan ly due o s a ch gela iniza ion (Zhou and The d hai 2007). 436 4 Conclusions 437 Changes p oduced du ing he e men a ion and baking o glu en- ee b eads depended 438 on he s uc u e and mo phology o s a ch g anules and lou pa icles. In gene al, 439 esul s showed ha he la ge and compac lou pa icles pa ially main ained hei 440 in eg i y du ing he kneading p ocess causing doughs o be mo e consis en and esis an 441 o shea s ess. This led o b eads wi h lowe olumes and ex u al p ope ies. On he 442 o he hand, he g anula mo phology, size, wa e abso p ion capaci y and pas ing 443 empe a u e a ec ed he way he s a ches in e ac ed. In his way, he bimodal size 444 dis ibu ion o whea s a ch was mo e p one o o m a uni o m con inuous s a ch-445 hyd ocolloid ma ix which was u he enhanced du ing baking as a consequence o he 446 low pas ing empe a u e o whea s a ch, en ailing ea lie amylose leaching. This led o 447 a dough wi h low consis ency bu high capaci y o e ain CO 2 du ing e men a ion, 448 esul ing in b eads wi h he highes speci ic olume and he bes ex u al pa ame e s 449 (low ha dness and high elas ici y, cohesi eness and esilience). These mechanis ic 450 ela ions be ween he de elopmen o he s a ch/ lou s uc u e, dough heology and 451 b ead quali y du ing b ead-making will p o ide use ul in o ma ion o he glu en- ee 452 b ead-making indus y. 453 454 5 Acknowledgemen s 455 The au ho s acknowledge he inancial suppo o he Spanish Minis y o Economy and 456 Compe i i eness (P ojec AGL2014-52928-C2) and he Eu opean Regional 457 De elopmen Fund (FEDER). 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CRC P ess, Boca Ra on, FL. 556 MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT MANUS CRIP T ACCEP TED ACCEPTED MANUSCRIPT Highligh s The heological e olu ion o glu en- ee doughs du ing e men a ion was s udied The ex u al e olu ion o glu en- ee b eads du ing baking was s udied Mechanis ic ela ions among s a ch, dough heology and b ead quali y we e ob ained Doughs wi h low consis ency and uni o m con inuous phase p o ided high olume b eads Whea s a ch was p one o o m a con inuous phase ha inc eased b ead quali y