Microwave radiation and protein addition modulate hydration, pasting and gel rheological characteristics of rice and potato starches

Accep ed Manusc ip Ti le: Mic owa e adia ion and p o ein addi ion modula e hyd a ion, pas ing and gel heological cha ac e is ics o ice and po a o s a ches Au ho s: Ma ina Villanue a, Bea iz De Lamo, Joanna Ha asym, Felicidad Ronda PII: S0144-8617(18)30950-0 DOI: h ps://doi.o g/10.1016/j.ca bpol.2018.08.052 Re e ence: CARP 13949 To appea in: Recei ed da e: 7-6-2018 Re ised da e: 6-8-2018 Accep ed da e: 11-8-2018 Please ci e his a icle as: Villanue a M, De Lamo B, Ha asym J, Ronda F, Mic owa e adia ion and p o ein addi ion modula e hyd a ion, pas ing and gel heological cha ac e is ics o ice and po a o s a ches, Ca bohyd a e Polyme s (2018), h ps://doi.o g/10.1016/j.ca bpol.2018.08.052 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. 1 Mic owa e adia ion and p o ein addi ion modula e hyd a ion, pas ing and gel heological cha ac e is ics o ice and po a o s a ches Ma ina Villanue a1, Bea iz De Lamo1, Joanna Ha asym1, 2, Felicidad Ronda1* 1 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. 2 Bio-Re Lab, Depa men o Bio echnology and Foods Analysis, Ins i u e o Chemis y and Food Technology, Facul y o Enginee ing and Economics, W ocław Uni e si y o Economics, W ocław, Poland. * Co esponding au ho : [email p o ec ed]; Tel: +34979108339; Fax: +34979108200 G aphical abs ac ACCEPTED MANUSCRIPT 2 Highligh s:  The swelling powe o po a o s a ch inc eased a e he mic owa e ea men  Mic owa e ea men dec eased he swelling powe o ice s a ch  The p o ein p esence changed he impac o mic owa es on s a ch p ope ies  Mic owa e adia ion al e ed he pas ing p ope ies o po a o s a ch  Mic owa e ea men inc eased iscoelas ic moduli o po a o s a ch gels Abs ac This s udy e alua ed o i s ime he e ec o Mic owa e (MW) adia ion on sys ems based on po a o and ice s a ches supplemen ed wi h 5% o calcium caseina e (CA) o soy p o ein isola e (SPI). The goal o his ea men was he physical modi ica ion o hese s a ch-based sys ems o p o ide ing edien s o new unc ionali ies. The hyd a ion and pas ing p ope ies as well as gel iscoelas ic ea u es we e e alua ed. Dynamic oscilla o y heological es s we e used. The e ec o MW ea men (MWT) depended on he s a ch bo anical o igin and was signi ican ly a ec ed by p o ein p esence and ype. MWT o s a ch+p o ein blends e ealed he mos no able changes when SPI was added. Adding i o ice s a ch dec eased swelling powe (-45%), al e ed iscome ic p o iles and ein o ced gel s uc u e wi h impo an inc eases in bo h iscoelas ic moduli (+160%-G’ and +58%-G’’). In blends wi h po a o s a ch, MWT inc eased wa e abso p ion capaci y (+115%), and dec eased wa e solubili y index (-82%). MWT o p o ein-po a o blends p omo ed gel s abili y, dec eased hei pas ing p o iles and esul ed in enhanced iscoelas ic moduli (+483-G’ and 243%-G’’). MWT combined wi h p o ein addi ion allows designing s a ch-based oods wi h ailo ed p ope ies. Keywo ds: Mic owa e; unc ional p ope ies; gel heology; po a o s a ch; ice s a ch 1. In oduc ion ACCEPTED MANUSCRIPT 3 The na i e o m o s a ch has limi ed applica ions in he ood indus y and is o en modi ied physically, chemically o enzyma ically o imp o e unc ional p ope ies. Physically modi ied s a ch is conside ed a na u al ma e ial and a highly sa e ing edien , much sa e and mo e p edic able han chemically modi ied s a ch. So a , he usage o physically modi ied s a ch in ood is no es ic ed by legisla ion, which is conside ed an ou s anding ad an age compa ed o chemically- o enzyma ically-modi ied s a ch applica ions (Klein e al., 2013). S a ch, as a mac o-cons i uen o many aw ma e ials and p ocessed oods, con ibu es o hei nu i ional and unc ional cha ac e is ics, as well as plays he pi o al ole as a ex u e modi ie (Walkens öm and He mansson, 1998) and in luences many quali y a ibu es o p ocessed oods (Chen e al., 2015). Rice and po a o s a ches a e among he mos commonly used o ex u e de elopmen , pa icula ly in glu en- ee (GF) p oduc s. Du ing mic owa e (MW) adia ion exposu e, ene gy is deli e ed di ec ly o he ma e ial, hea ing he en i e sample in bulk a a as e hea ing a e han con en ional hea ing (Bilbao-Sáinz e al., 2007). The e o e, MW adia ion-induced hea ing is sui able o modi ying s a ches physically and imp o ing hei unc ional p ope ies. In mois ened s a ches, he in e molecula s uc u e changes om MW-deli e ed ene gy lead o changes in wa e abso p ion abili y, solubili y and swelling powe , as well as in s a ch gela iniza ion, syne esis and pas e iscosi y (B aso eanu and Nem anu, 2014). The e a e many ac o s ela ed o bo h s a ch cha ac e is ics (bo anical o igin, wa e con en , densi y, dielec ic p ope ies, e c.) and mic owa e i adia ion condi ions ( equency, powe and exposu e ime) ha signi ican ly a ec how g anula s a ch esponds o MW ea men (MWT) (B aso eanu and Nem anu, 2014). Za a eze e al. (2010) showed ha mic owa e i adia ion a ec ed he pas ing p ope ies o high- amylose ice s a ches mo e in ensely han in hose wi h lowe amylose con en . ACCEPTED MANUSCRIPT 4 Ve meylen e al. (2006) concluded ha highe ea men empe a u es and mois u e con en s gene ally caused he la ges changes in s a ches. As applied empe a u e and mic owa e powe le el inc eased, he peak iscosi y o po a o s a ch dec eased (Nadiah e al., 2015), while he gela iniza ion empe a u e inc eased (Nadiah e al., 2015; Ve meylen e al., 2006). Villanue a e al. (2018a) concluded ha he MW adia ion abso p ion capaci y o ice lou exhibi ed a posi i e quad a ic ela ionship wi h wa e con en , while he lou d y ma e did no show any adia ion abso p ion capaci y. They also obse ed changes in physical p ope ies o ea ed ice lou , such as ice lou pa icle mo phology o he c ys allini y/amo phous egion a io. Beyond physico-chemical and unc ional conside a ions, s a chy glu en- ee oods a e o en conside ed nu i ionally poo compa ed o whea -based coun e pa s due o hei de icien p o ein/s a ch a io (Villanue a e al., 2015). The addi ion o p o ein is a iable al e na i e o imp o e he nu i ional p ope ies o hese p oduc s. P o eins a e also known o modi y s uc u e and ex u e in oods (Villanue a e al., 2018b). Soy p o eins isola e (SPI) and calcium caseina e (CA) ha e been in es iga ed in GF applica ions because o hei e ec on nu i ion and s uc u e o p oduc s (K upa-Kozak e al., 2011; Ronda e al., 2014). S a ch-p o ein in e ac ions a ec he heological, pas ing, gela iniza ion, ex u al and physicochemical p ope ies o ood sys ems (Gallaghe e al., 2003; Ronda e al., 2011; Ronda e al., 2014; Villanue a e al., 2015). As a as we know, how MWT a ec s p o ein-s a ch mix u es has no been s udied so a , in spi e o how impo an endogenous p o eins seem o be in he e ec o hea -mois u e ea men (HMT) on ea ed- lou p ope ies (Puncha-a non and U apap, 2013). As p o ein and s a ch blends can se e as model sys ems o he s udy o mo e complex ma ices such as lou , he impac o hei in e ac ions on changes appea ing in MW ea ed samples needs o be be e unde s ood. The main objec i e o his s udy was o in es iga e he ACCEPTED MANUSCRIPT 5 combined e ec o mic owa e adia ion and p o ein addi ion (5% CA o SPI) on he hyd a ion, pas ing p ope ies and gel iscoelas ic p ope ies o po a o and ice s a ches.. The ul ima e goal o his s udy is o p o ide s a ch-based ing edien s wi h ailo ed unc ional p ope ies. 2. Ma e ial and me hods 2.1. Ma e ials Rice s a ch and po a o s a ch we e supplied by Fe e Alimen ación S.A. (Ba celona, Spain). Soybean p o ein isola e (SPI) Sup o 500-E IP (pu i y ~ 90%) was ob ained om P o eedo a hispano-holandesa S.A. (Ba celona, Spain), and calcium caseina e (CA) (pu i y ~ 96%) om A mo P o éines (Sain -B ice-en-Coglès, F ance). P o ein added in all he s a ch+p o ein blends was 5 g/100 g blend. Twel e di e en samples we e in es iga ed: na i e and MW- ea ed ice and po a o s a ches alone and blended wi h 5 g/100 g o CA o SPI p o eins. 2.2. Mic owa e ea men o s a ch samples Samples we e ea ed by MW adia ion a ini ial mois u e con en o 30%. The l mois u e con en o he samples was measu ed ollowing he Ame ican Associa ion o Ce eal Chemis s (AACC) 44–19 me hod and he wa e necessa y o adjus o 30% mois u e con en was sp ayed on o he samples while hey we e mixed in a Bea Teddy Mixe (Bea 5L Teddy, Swansea, UK) o 15 min. Samples we e hen he me ically sealed in bags and held 24 h a 4 ± 2°C o he mois u e equilib a ion. Hyd a ed samples (100.0 g each) we e hea ed in a SHARP R-342 (Osaka, Japan) mic owa e o en in a cylind ical polye hylene con aine closed wi h needle-punched, mic owa e-sa e ood g ade ilm. The homogenous dis ibu ion o adia ion on he sample is o c ucial impo ance. So, he con aine was s i ed cons an ly o ensu e a uni o m ene gy and empe a u e dis ibu ion du ing ea men as desc ibed in ACCEPTED MANUSCRIPT 6 Villanue a e al. (2018b). MW adia ion equency was 2450 MHz and he powe , 900W. MW was applied in cycles o 20 s adia ion wi h 40 s down ime, o a o al o 32 min and 640 s o MWT; a e wa ds, samples we e le o cool o 5 minu es. Agglome a es o med du ing ea men we e manually disin eg a ed in a labo a o y mo a o < 0.5 mm. Sample empe a u e du ing MWT was measu ed wi h Tes o e m he mome e s ips o di e en scales, and 0.5ºC accu acy (Ins umen os Tes o S.A., Ba celona, Spain), as desc ibed in Villanue a e al. (2018a). Each measu emen was made in duplica e. The empe a u e e olu ion cu es ob ained o he po a o and ice s a ch sys ems s udied we e equi alen o hose epo ed o 30% mois ened- ice lou in ou p e ious wo k (Villanue a e al., 2018a). The empe a u e eached and main ained a e 4 min o ea men was 157±5ºC o all samples. 2.3. Amylose con en The amylose con en o he s a ch samples was de e mined using a colo ime ic assay ki o amylose/amylopec in a io de e mina ion in s a ch (Megazyme In e na ional I eland L d., I eland), acco ding o he manu ac u e ’s p ocedu e based on Gibson e al., (1997) me hod. The me hod de e mines he soluble amylose in ace a e/sal solu ion suscep ible o α-amylase/ amyloglucosidase diges ion. Each sample was analysed a leas in duplica e. 2.4. Hyd a ion p ope ies Wa e abso p ion capaci y (WAC) o he samples was de e mined by he cen i uga ion me hod desc ibed by Abebe e al., (2015). Two g ams o sample (ws) we e mixed wi h 20 mL o dis illed wa e in 50 mL cen i uge ubes. The dispe sions we e hold a oom empe a u e o 30 min wi h occasionally o exed (Heidolph Reax, Schwabach, Ge many) and ollowed by cen i uga ion o 30 min a 3000×g (The mo Fishe Scien i ic, Wal ham, USA). The supe na an was emo ed and weighed (ws+w) and ACCEPTED MANUSCRIPT 7 esul s we e exp essed as g ams o wa e e ained pe g am o sample. Wa e abso p ion index (WAI) and wa e solubili y index (WSI) we e measu ed wi h sligh modi ica ion o he me hod used by Abebe e al. (2015). Each sample o 2.5 g (w0) was dispe sed in 30 ml o dis illed wa e in a ed cen i uge ubes. A e cooking o 10 min in a 90ºC wa e ba h, samples we e cooled o oom empe a u e and cen i uged a 4000xg o 10 min. The supe na an was pou ed in o a p e-weighed e apo a ing capsule o de e mine he solid con en and he sedimen was weighed (wss). The weigh o soluble solids was eco e ed by e apo a ing he supe na an o e nigh a 110ºC (wds). WAC, WAI, WSI and swelling powe (SP) we e calcula ed using he ollowing equa ions: 𝑊𝐴𝐶(𝑔 𝑔) = 𝑤𝑠+𝑤−𝑤𝑠 𝑤𝑠 (1) 𝑊𝐴𝐼 (𝑔 𝑔) = 𝑤𝑠𝑠 𝑤0 (2) 𝑊𝑆𝐼( 𝑔 100𝑔) = 𝑤𝑑𝑠 𝑤0×100 (3) 𝑆𝑃(𝑔 𝑔) = 𝑤𝑠𝑠 (𝑤0−𝑤𝑑𝑠) (4) All esul s we e e e ed o d y ma e o a oid he e ec o di e en wa e con en in he samples. 2.5. The mo iscous es : Viscome ic p o ile Following AACC In e na ional Me hod 76-21.01 S anda d 2, iscome ic p o iles o MW- ea ed and un ea ed samples we e ob ained using a Kinexus P o+ heome e (Mal e n Ins umen s L d., Mal e n, UK) supplied wi h s a ch pas ing cell and con olled by Space so wa e. Samples o ice s a ch (3 g, 14% mois u e-based) o po a o s a ch (2 g, 14% mois u e-based) wi h o wi hou added p o ein we e ans e ed o a canis e whe e 25 mL±0.1 mL o dis illed wa e was added. Each s a ch suspension was equilib a ed a 50 ºC o 1 min, hea ed o 95 ºC a a a e o 6 ºC/min, main ained a 95 ºC o 5 min, hen cooled o 50 ºC a a a e o 6 ºC/min, and main ained a 50 ºC o 2 ACCEPTED MANUSCRIPT 8 min. Paddle speed was se a 960 pm o he i s 10 s and hen 160 pm o he es o he analysis. Each sample was analyzed a leas in duplica e. Pa ame e s calcula ed om he pas ing p o iles we e pas ing empe a u e (PT), peak iscosi y (PV), ough iscosi y (TV), b eakdown (BD), inal iscosi y (FV), se back (ST) and peak ime. 2.6. Rheological measu emen s Dynamic oscilla o y es s o he wel e po a o and ice s a ch gels we e pe o med wi h Kinexus P o+ heome e (Mal e n Ins umen s L d., Mal e n, UK) wi h pa allel pla e geome y (40 mm diame e ) o se a ed su ace and wi h 1 mm o wo king gap. The gel samples we e made ollowing he same p ocedu e desc ibed o he mo iscous es s. Di e en concen a ions o ice and po a o s a ches we e used o p epa e hei gels, so ha he gels p oduced om na i e s a ches we e o simila consis encies. Jus a he end o he pas ing es he gel was emo ed om he canis e and placed be ween he pla es, he sample excess was emo ed and he sample was le o es o 5 min o allow elaxa ion. Tempe a u e was s abilized a 25ºC wi h a Pel ie pla e con olle . S ess sweeps we e pe o med om 0.1 o 500 Pa a a cons an 1 Hz equency. F equency sweeps we e ca ied ou om 10 o 1 Hz in he linea iscoelas ic egion (a a cons an alue o 1 Pa). F equency sweep da a we e i ed o po en ial equa ions as desc ibed by Ronda e al. (2014). All gels we e p epa ed a leas in duplica e and heological es s we e also pe o med in duplica e. 2.7. S a is ical analysis S a g aphics Cen u ion .6 (Bi s eam, Camb idge, MN, USA) was used o mul i ac o analysis o a iance (ANOVA) o he da a. The Leas Signi ican Di e ence es was used o e alua e signi ican di e ences (p < 0.05) be ween samples. 3. Resul s and discussion 3.1. Amylose con en ACCEPTED MANUSCRIPT 15 high humidi y hea ing pe iod wi h a d y hea ing pe iod could explain he in e media e beha iou ha we obse ed in ice s a ch wi h espec o ha ob ained by Puncha-a non and U apap (2013) and Qiu e al. (2016). ACCEPTED MANUSCRIPT 16 Table 2. E ec o mic owa e ea men and p o ein p esence on he pas ing p ope ies o s a ch samples. S a ch P o ein MW ea men PT(ºC) PV (Pa·s) TV ( Pa·s ) BD ( Pa·s ) FV ( Pa·s ) ST ( Pa·s ) Peak ime (s) Rice 0 0 74.6 ab 2.55 c 1.25 c 1.30 b 2.24 c 0.99 d 665 d 1 73.9 a 2.73 d 1.23 c 1.49 c 2.29 c 1.06 e 627 ab CA 0 75.8 b 2.09 a 1.02 a 1.08 a 1.82 a 0.80 b 650 c 1 74.6 ab 2.27 b 1.11 b 1.16 a 2.00 b 0.88 c 635 b SPI 0 73.9 a 2.32 b 1.03 a 1.30 b 1.72 a 0.69 a 618 a 1 74.3 a 2.68 cd 1.35 d 1.33 b 2.31 c 0.96 d 620 a Analysis o a iance and signi icance (p- alues) Fac o 1 (p o ein ype) * *** *** *** *** *** *** Fac o 2 (MW ea men ) ns *** *** ** *** *** *** Fac o 1x2 ns ns *** ns *** *** ** Po a o 0 0 70.0 a 6.88 e 2.06 b 4.82 d 2.84 ab 0.78 a 358 a 1 76.2 b 1.85 b 2.11 b 0.00 a 3.70 b 1.59 b 687 c CA 0 71.4 a 3.15 d 1.66 a 1.49 c 2.34 a 0.67 a 518 b 1 79.0 c 1.60 a 1.59 a 0.00 a 2.43 a 0.84 a 860 d SPI 0 70.5 a 2.72 c 1.50 a 1.22 b 2.39 a 0.89 ab 538 b 1 78.1 c 1.92 b 2.01 b 0.00 a 3.29 ab 1.28 ab 762 cd Analysis o a iance and signi icance (p- alues) Fac o 1 (p o ein ype) ** *** ** *** * ns ** Fac o 2 (MW ea men ) *** *** * *** * * *** Fac o 1x2 ns *** ** *** ns ns ns P o ein: 0: wi hou p o ein; CA: 5% calcium caseina e; SPI: 5% soy p o ein isola e. MW ea men : 0: wi hou ea men ; 1: wi h ea men . PT: pas ing empe a u e, PV: peak iscosi y, TV: ough iscosi y, BD: b eakdown, FV: inal iscosi y, ST: se back. The di e en le e s in he co esponding column wi hin each s a ch ype indica e s a is ically signi ican di e ences be ween means a p<0.05. Analysis o a iance and signi icance: *** p<0.001. ** p<0.01. * p<0.05. ns: no signi ican . ACCEPTED MANUSCRIPT 17 0 10 20 30 40 50 60 70 80 90 100 0 0,5 1 1,5 2 2,5 3 0 200 400 600 800 1000 1200 1400 Tempe a u e (ºC) Viscosi y (Pa·s) Time (s) 0 10 20 30 40 50 60 70 80 90 100 0 1 2 3 4 5 6 7 8 0 200 400 600 800 1000 1200 1400 Tempe a u e (ºC) Viscosi y (Pa·s) Time (s) Figu e 1. Pas ing p o iles o na i e and modi ied samples o ice s a ch (A) and po a o s a ch (B). S a ch wi hou p o ein a e ep esen ed by , wi h 5% calcium caseina e by , and wi h 5% soy p o ein isola e by . Mic owa e- ea ed p o ein- ee samples a e ep esen ed by , mic owa e- ea ed samples wi h 5% calcium caseina e by and wi h 5% soy p o ein isola e by . The empe a u e p o ile is ep esen ed by in he second axis. B) A) ACCEPTED MANUSCRIPT 18 The low peak ime o na i e po a o s a ch inc eased signi ican ly wi h MWT, eaching he peak alues o ice s a ch. Na i e po a o s a ch showed a much highe BD han ice s a ch, 4.82 e sus 1.30 Pa·s (Table 2). A low BD alue means high pas e s abili y e sus hea and shea , and is ela ed o g anule igidi y and high lipid con en (Singh e al., 2003). Adding p o ein inc eased he pas e s abili y o po a o s a ch, dec easing he BD alue by 69% and 75% o CA and SPI, espec i ely. Howe e , in he case o ice s a ch, only CA had an e ec and i was much sligh e . MWT ha dly a ec ed he BD alue o he ice s a ch samples. Howe e , he ea men did educe po a o s a ch BD o ze o, ega dless o he p esence o p o eins. This e lec s he abili y o MWT o inc ease po a o s a ch gel s abili y e sus hea ing and shea ing. This beha iou has al eady been obse ed in ube s a ches (po a o and apioca) when hey we e mois ened and he mally modi ied ei he by con ec ion hea ans e (Klein e al., 2013) o MW adia ion (Nadiah e al., 2015). ANOVA analysis (Table 2) e ealed ha p o ein ype and MW ea men had a singula ly s ong e ec on PT, PV and BD alues o ice s a ch, wi hou any mu ual in e ac ion o hese ac o s. Howe e , po a o s a ch was highly suscep ible, no only o p o ein ype supplemen a ion o MWT bu also o he double in e ac ion o he wo ac o s on PV and BD. The inal iscosi y (FV) o he gels o med a e subsequen cooling dec eased in p esence o ei he animal o ege al p o ein. The same had p e iously been epo ed by Ronda e al. (2014). The e ec was mo e p onounced o ice s a ch gels han o po a o gels. MWT inc eased he FV o ice+p o ein gel, while po a o s a ch FV was una ec ed. ANOVA analysis showed ha each o he main ac o s in luenced inal iscosi y, bu he double e ec was ound only in ice s a ch. Se back iscosi y (ST), mainly ela ed o he amylose leached om s a ch g anules and hei endency o eo ganise a e gela inisa ion (Miles e al., 1985), was equi alen o bo h na i e s a ches in ou s udy. Adding p o eins signi ican ly (p<0.001) dec eased his ACCEPTED MANUSCRIPT 19 alue in ice s a ch blends, while po a o alues emained una ec ed. MWT inc eased ST in all ice s a ch-based o mula ion and p o ein- ee po a o s a ch. In he case o po a o s a ch he inc ease was om 0.78 o 1.59 Pa·s. The ANOVA esul s con i med ha he simple and double- ac o e ec s on ice s a ch ST we e highly signi ican (p < 0.001). Howe e , excep o MWT, he e we e almos no e ec s on po a o s a ch. 3.4. Gel heological p ope ies The iscoelas ic p ope ies o gel samples we e s udied by dynamic oscilla o y es s, applying a sinusoidal s ess o he samples. The ob ained iscoelas ic moduli as a unc ion o s ess a e shown in Figu e 2. The pa ame e s ob ained om i ing he mechanical spec a o powe law a e summa ized in Table 3. The high R2 alues ob ained, always abo e 0.998 o all samples, demons a e ha he sys ems s udied adjus ed well o he powe law model. S ess sweeps made i possible o es ablish he linea iscoelas ic egion (LVR) by iden i ying he maximum s ess (τmax) ha samples could ole a e conse ing hei s uc u e. These alues a e also included in Table 3. The elas ic modulus, G’, o gels made om un ea ed s a ches s a ed o dec ease a 108 and 267 Pa o ice and po a o s a ch, espec i ely. This indica ed ha s a ch sample s uc u es had di e en esis ances o s ess-induced up u e. Acco ding o Villanue a e al. (2018b), po a o gels p ese ed hei s uc u e beyond ice s a ch gels, meaning ha po a o gels had good mechanical esis ance and ha dness. Adding p o eins o ice s a ch gels educed hei esis ance o b eakage and led o alues o τmax 36% and 49% lowe han hose o p o ein- ee gels o CA and SPI, espec i ely. The e ec on po a o s a ch gels di e ed depending on p o ein ype: CA dec eased τmax by 19%, while SPI inc eased i by 14%, compa ed o p o ein- ee po a o gels. MWT inc eased he s eng h o ice gels, al hough his e ec was only signi ican in SPI-en iched ice gels. The e ec o MW adia ion on po a o gels was he opposi e and s onge , being much mo e ACCEPTED MANUSCRIPT 20 ema kable in he p esence o p o eins: τmax dec eased by 60% in gels wi hou p o ein, 88% in CA p o ein-en iched gels and 72% in SPI-en iched gels. ACCEPTED MANUSCRIPT 21 Figu e 2. S ain sweeps o ice (A) and po a o (B) s a ch gels wi hou p o ein addi ion (A1 and B1), wi h 5% calcium caseina e (A2 and B2) o 5% soy p o ein isola e (A3 and B3). Non- ea ed samples a e ep esen ed wi h ci cles and MW- ea ed samples wi h iangles. The elas ic modulus, G’, is ep esen ed using solid symbols and he iscous modulus, G’’, wi h open symbols. Rice A1 1 10 100 1000 0,1 110 100 1000 G', G'' (Pa) S ess (Pa) 1 10 100 1000 0,1 1 10 100 1000 G', G'' (Pa) S ess (Pa) Po a o B1 B2 A3 A2 B3 ACCEPTED MANUSCRIPT 22 Table 3. E ec o mic owa e ea men and p o ein p esence on he dynamic heological cha ac e is ics o s a ch gels. S a ch P o ein MW ea men G’1 (Pa) a G’’1 (Pa) b ( an δ)1 c max (Pa) Rice 0 0 52 ab 0.17 b 14 ab 0.36 a 0.27 bc 0.18 ab 108 c 1 74 bc 0.15 a 17 ab 0.35 a 0.23 ab 0.20 b 119 c CA 0 41 a 0.18 b 12 a 0.36 a 0.29 c 0.18 a 69 ab 1 47 a 0.17 b 13 ab 0.35 a 0.27 bc 0.18 ab 85 b SPI 0 39 a 0.19 b 12 a 0.36 a 0.30 c 0.17 a 55 a 1 88 c 0.14 a 19 b 0.35 a 0.22 a 0.20 b 105 c Analysis o a iance and signi icance (p- alues) Fac o 1 (p o ein ype) ns ns ns ns ns ns ** Fac o 2 (MW ea men ) ** ** * ns ** * ** Fac o 1x2 ns ns ns ns ns ns * Po a o 0 0 53 a 0.181 e 16 a 0.29 b 0.31 e 0.11 ab 267 d 1 421 d 0.077 a 51 b 0.22 a 0.12 a 0.15 c 101 b CA 0 46 a 0.184 e 14 a 0.28 b 0.31 e 0.10 a 216 c 1 171 b 0.141 c 39 b 0.28 b 0.23 c 0.14 bc 26 a SPI 0 53 a 0.164 d 14 a 0.28 b 0.27 d 0.12 abc 304 d 1 309 c 0.099 b 48 b 0.23 a 0.15 b 0.14 bc 84 b Analysis o a iance and signi icance (p- alues) Fac o 1 (p o ein ype) *** *** ns * *** ns ** Fac o 2 (MW ea men ) *** *** *** ** *** ** *** Fac o 1x2 *** *** ns * *** ns ns P o ein: 0: wi hou p o ein; CA: 5% calcium caseina e; SPI: 5% soy p o ein isola e. MW ea men : 0: wi hou ea men ; 1: wi h ea men . G’1, G’’1, and ( an δ)1, ep esen he elas ic and iscous moduli and he loss angen a a equency o 1 Hz. The a, b and c exponen s quan i y he dependence deg ee o dynamic moduli and he loss angen wi h he oscilla ion equency, ω. τmax ep esen s he maximum s ess ha GF ma ices can ole a e in he LVR. The di e en le e s in he co esponding column wi hin each s a ch ype indica e s a is ically signi ican di e ences be ween means a p<0.05. Analysis o a iance and signi icance: *** p<0.001. ** p<0.01. * p<0.05. ns: no signi ican . ACCEPTED MANUSCRIPT 23 In all samples, G’ and G” inc eased wi h equency, G’ being highe han G”, which indica ed ha elas ic cha ac e p e ailed o e iscous ac o (Xie e al., 2013). The gel iscoelas ic moduli G1’ and G1’’ and he loss angen a 1 Hz we e he same o he wo na i e s a ches (po a o and ice) ega dless o p o ein p esence o ype. Howe e , MWT always inc eased gel consis ency, his impac was dependen on p o ein p esence and ype and s a ch ype. The e ec o MWT on gel heological p ope ies was also s ongly dependen on he bo anical o igin o he s a ches. Bo h iscoelas ic moduli inc eased in MW- ea ed po a o s a ch samples, while only SPI addi ion a ec ed he moduli in ea ed ice ones. The G1’ and G1’’ inc eases om MWT o po a o gels we e ela ed o he gela iniza ion le el and he ne wo k de elopmen o chains leached om he s a ch g anules du ing he MW ea men (Xie e al., 2013). As a consequence o MWT, he inc ease in he elas ic modulus was always mo e no iceable han in he iscous modulus. Consequen ly, MWT was always accompanied by a d op in he loss angen , which mean a ein o cemen o he elas ic cha ac e o gels made om MW- ea ed s a ches o s a ch+p o ein blends compa ed o he un ea ed coun e pa s. Simila esul s ha e been epo ed o he mally-modi ied po a o s a ch (G yszkin e al., 2014). Tan δ alue was e y low o all samples (always < 0.4), which co esponded wi h a well c oss-linked ne wo k. Adding SPI dec eased he gel loss angen o po a o s a ch, bu ice gels we e una ec ed by p o ein addi ion. Villanue a e al. (2018b) epo ed ha he heological p ope ies o ice and po a o s a ch gels o i ied wi h egg albumen o SPI depended signi ican ly on he double in e ac ion (s a ch x p o ein), concluding ha he same p o ein exe ed a di e en e ec depending on he s a ch sou ce. The alues o he “a” exponen we e always below he exponen “b”. This mean ha G’’ inc eased wi h equency as e han G’. This yielded an inc ease in an  wi h equency, which could also be con i med om he posi i e alue o exponen “c” (Table 3). Consequen ly, gels inc eased hei iscous beha iou wi h equency. ACCEPTED MANUSCRIPT 24 4. Conclusions MWT has been shown o be success ul in modi ying s a ches and hei p o ein mix u es and has p o en o be e icien in al e ing he physical p ope ies o p oduc s made om hem. I is pa icula ly use ul in he de elopmen o p o ein-en iched GF p oduc s. The esul s showed ha he e ec o MWT is dependen on s a ch sou ce and p o ein ype. MWT changed he hyd a ion p ope ies and enhanced wa e abso p ion index and swelling powe in po a o samples, while i dec eased hem in ice s a ch samples, ega dless o ype o p o ein added. MW adia ion in luenced mainly he pas ing p ope ies o po a o s a ch, inc easing pas ing empe a u e and se back, and dec easing peak iscosi y and b eakdown. The ea men inc eased he iscoelas ic moduli signi ican ly and dec eased he loss angen o po a o gels bo h wi h and wi hou p o eins. In gene al, he inclusion o p o eins inc eased WAC, WAI, SP and WSI, dec eased he iscosi y o gels and inc eased hei s abili y, being he e ec mo e ma ked o SPI inco po a ion. This esea ch helps unde s and he changes ha occu du ing MW ea men s o model sys ems and can help design and imp o e he quali y o new p oduc s. MW-assis ed hea ing is an inno a i e lou ea men me hod and can be used as an al e na i e o chemical modi ica ion. Acknowledgemen s The au ho s a e g a e ul o he inancial suppo o he Spanish Minis e io de Economía y Compe i i idad and he Eu opean Regional De elopmen Fund (FEDER) (AGL2015- 63849-C2-2-R) and o he Jun a de Cas illa y León/FEDER VA072P17. Joanna Ha asym (MSCA) hanks he Eu opean Commission o he Indi idual Fellowship ACCEPTED MANUSCRIPT