Vliv přídavku karagenanů na viskoelastické vlastnosti modelových tavených sýrů

ACTA UNIVERSITATIS AGRICULTURAE ET SILVICULTURAE MENDELIANAE BRUNENSIS SBORNiK MENDELOVY ZEMEDELSKE A LESNICKE UNIVERZITY V BRNE RocnikLV 7 Cislo 5, 2007 THE EFFECT OF ADDITION OF SELECTED CARRAGEENANS ON VISCOELASTIC PROPERTIES OF MODEL PROCESSED CHEESE SPREADS M. Cemiko a, F. Buiika, V. Pa linek, P. B ezina, J. H abe, S. K acma Recei ed: May 20,2007 Abs ac CERNIKOV A, M., BuNKA, F., PAVLlNEK, ., BREZINA, P., HRABE, J., KRACMAR, S.: The e ec o addi ion o selec ed ca ageenans on iscoelas ic p ope ies o model p ocessed cheese sp eads. Ac a uni . ag ic. e sil ic. Mendel. B un., 2007, LV, No.5, pp. 51-58 The e ec o 0.25% w/w K-ca ageenan and -ca ageenan on iscoelas ic p ope ies o p ocessed chee- se we e s udied using model samples con aining 40% w/w d y ma e and 45 and 50% w/w a in d y ma e . Expe imen al samples o p ocessed cheese we e e alua ed a e 14 days o s o age a he em- pe a u e o 6 ± 2°C. Basic pa ame e s o p ocessed cheese samples unde s udy (i.e. hei d y ma - e con en and pH) we e no di e en (P ;::: 0.05). The e we e no s a is ically signi ican di e ences in alues o s o age modulus G' [Pal, loss modulus G" [Pal and angen o phase shi angle an () [-] o he e e ence equency o 1 Hz be ween p ocessed cheese wi h K-ca ageenan applied in he o m o powde and in he o m o aqueous dispe sion (P;::: 0.05). The addi ion o 0.25% w/w K-ca ageenan and -ca ageenan (in he powde o m) esul ed in an inc ease in s o age (G') and loss (G") moduli and a dec ease in alues o an () (P < 0.05). As compa ed wi h con ol (i.e. wi hou added ca ageenans), sam- ples o p ocessed cheese became i me . Io a-ca ageenan added in he powde o m in concen a ion o 0.25% w/w showed a mo e in ensi e e ec on he inc ease in i mness o p ocessed cheese unde s udy han K-ca ageenan (P < 0.05). p ocessed cheese, a in d y ma e , kappa-ca ageenan, io a-ca ageenan, heology Hyd ocolloids (ca ageenan, pec in, modi ied s a ch, gums, e c.) a e used as s abilizing and hickening agen s in he p ocessed cheese p oduc ion. P ocessed cheese is cheese-base ood p oduced by comminu- ing, mel ing and emulsi ying in o a smoo h homoge- neous mol en blend, one o mo e na u al cheeses and op ional ing edien s using hea , mechanical shea and emulsi ying sal s (Guinee e al., 2004). Ca ageenans a e anionic linea polysaccha ides ex ac ed om ed seaweed (Rhodophyceae), consi- s ing o al e na ing a-1,4 and ~-1,3 linked anhyd o- galac ose esidues. The e a e h ee majo ac ions (K- kappa, -io a and A-lambda) wi h a ying numbe and posi ion o sulpha e g oups. Kappa-ca ageenan and -ca ageenan unde go a empe a u e-dependen coil 51 (diso de ed s a e) o helix (o de ed s a e) ansi ion (in aqueous solu ion). Kappa-ca ageenan usually o ms i m, b i le gels and -ca ageenan usually gene a es so elas ic gels. Gel s eng h o bo h polysaccha ides s ongly depends on ca ions p esen -K-ca ageenan is especially sensi i e o po assium and -ca ageenan o calcium. Lambda-ca ageenan is no able o build up s able gels (Sy be e aI., 1998; Tziboula and Ho ne, 1999; Imeson, 2000; Singh e aI., 2003; Ribei o e al., 2004; Spagnuolo e al., 2005). Casein micelles a e complex o indi idual casein ac ions: as!' aS2' ~ and K-casein. Kappa-casein is loca ed on he pe iphe y o micelle, p o iding an elec os a ic s abilising ou e laye (Bou io e aI., 1999; de K ui and Tuinie , 2001). In e ac ions ----------- 52 M Ce n kowi, F Buiika, V. Pa l nek, P. B ezina, J. H abe, S. K acma K-ca ageenan and -ca ageenan wi h milk p o eins, especially wi h casein micelles, ha e been widely s u- died (Thaiudom and Go , 2003). "Milk eac i i y" o K-ca ageenan and -ca ageenan is mos ly a ibu ed o in e ac ions be ween he nega i e sulpha ed g oups o he ca ageenans and a posi i ely cha ged egion si ua ed be ween esidues 97 and 112 o K-casein a he su ace o he casein micelle (Gamie e al., 2003; Vega e al., 2005). Beha iou o milk p o ein-ca a- geenan sys ems depends on a many pa ame e s, such as hyd ocolloid, p o ein and dispe sed pa icle con- cen a ion, pH, ionic en i onmen (e.g. po assium and calcium concen a ion), suga con en , empe a u e, molecula weigh , milk p o ein p ocessing, he mal his o y (Sy be e al., 1998). In na u e cheese ( aw ma e ial o p ocessed cheese p oduc ion) he e is nei he casein in micella o m no K-casein ("na u al" wi h ull 169 esidues). In he p ocessed cheese he e is no signi ican amoun o micella casein (excluding o si ua ion when skim milk powde o o he ma e ials con aining casein micelles a e added). Only a ew s udies ha e deal wi h in e ac ions o K-ca ageenan and -ca ageenan wi h non-micella casein sys ems o indi idual casein ac ions excep K-casein (e.g. as and P). Lynch and Mul ihill (1996) p oposed ha abili y o as and P- caseins o in e ac wi h ca ageenans equi es he p e- sence o calcium ions and es e -bound phospho us on se yl esidues o casein ac ions. The objec o his pape was o use K-ca ageenan and -ca ageenan in p ocessed cheese p oduc ion (p oduc s wi h 45% w/w and 50% w/w a in d y ma - e ) and ind ou di e ences in iscoelas ic p ope ies o bo h o applied ca ageenans in model concen a- ion. MATERIAL AND METHODS Model samples o p ocessed cheese sp eads con ai- ning 40% w/w d y ma e (DM) and 45% w/w o 50% w/w a in d y ma e (FDM) we e manu ac u ed om a mix u e o na u al cheese (Edam block cheese wi h 30% w/w FDM and Edam block cheese wi h 45% w/ w FDM), bu e , deionized wa e ( o assu e a cons an concen a ion o ions) and comme cially supplied emulsi ying sal s. In he i s pa o his expe imen (G oup I), he e - ec o he o m o added K-ca ageenan on iscoelas- ic p ope ies o model p ocessed cheeses con aining 45% w/w and 50% w/w FDM was s udied. Kappa- ca ageenan was applied in o bo h ypes o cheese (i.e. wi h 45% w/w and 50% w/w FDM) in concen a- ion o 0.25% w/w, pa ly in he o m o powde and pa ly in he o m o aqueous dispe sion (in deionized wa e , which was used in such amoun ha did no in luenced he DM con en in he inal p oduc s). Two con ol samples ( o bo h FDM con en s unde s udy) we e p epa ed as well. In he second pa o his expe imen , ou g oups o model p ocessed cheese samples we e used (i = II, III, IV, V). All o hem we e manu ac u ed using he same echnology (see below), and ca ageenans in powde o m we e applied o each o hem. In pa icula , each o hese ou g oups consis ed o six ypes o sam- ples (ba ches): (1) p oduc wi h 0.25% w/w o K-ca - ageenan (Ke) and wi h 45% w/w FDM; (2) p oduc wi h 0.25% w/w o -ca ageenan (IC) and wi h 45% w/w FDM; (3) p oduc wi h 0.25% w/w KC and 50% w/w FDM; (4) p oduc wi h 0.25% w/w IC and 50% w/w FDM; (5) con ol sample wi hou ca ageenan and wi h 45% w/w FDM; (6) con ol sample wi hou ca ageenan and wi h 50% w/w FDM. In he second pa o his expe imen , samples o na u al cheese wi h a di e en deg ee o ma u i y we e used (G oup II - 6 weeks; G oup III - 8 weeks; G oup IV - 10 weeks and G oup V - 16 weeks). In indi idual es g oups, he ini ial i mness o con ol samples was di e en (see alues o s o age (GO) and loss (G") moduli in Tabs II and III). Fo ha eason he conclusions con- ce ning e ec s o KC and IC addi ion we e ela ed o he con ol sample o each g oup. This expe imen al layou was used o each a highe in o ma ion capa- bili y and ep esen a i eness o esul s o indus ial p ac ice whe e i is some imes di icul o assu e a cons an deg ee o ma u i y o he basic aw ma e ial (i.e. na u al cheese). P ocessed cheese sp eads we e manu ac u ed using he equipmen Vo we k The momix TM 21 blende cooke (Vo we k & Co. The momix; GmbH, Wup- pe al, Ge many); he mel ing empe a u e was 92 °C ( he ime in e al necessa y o each he mel ing em- pe a u e was 9-10 min. and he mel ing empe a u e was main ained o 1 min.). The ea e he p ocessed cheese was cooled down o 6 ± 2 °C and s o ed o 14 days a he same empe a u e. Ca ageenans (KC and IC) used in his expe imen we e supplied by he com- pany Sigma Ald ich, Inc., S . Louis, USA. The con en o DM was de e mined a he empe- a u e o 102 ± 1 DC, which was main ained ill he loss o weigh emained o be cons an (Be ge e al., 2002). Values o pH we e measu ed wi h he pH- me e G y 209 S wi h a combined glass elec ode a 22 ± 1 DC. Viscoelas ic p ope ies o p ocessed cheese samples we e measu ed by Bohlin Gemini (Mal e n Ins umen s, UK) heome e wi h pa allel pla e geo- me y (diame e o 40 mm, gap 1 mm) a he empe- a u e o 20 DC. All he expe imen s we e pe o med in he con ol shea s ess mode a equency anging om 0.1 o 50 Hz. The ampli ude o shea s ess 50 Pa was chosen in he egion o linea iscoelas ici y. The exposed edge o pa allel pla es geome y was co e ed wi h a hin laye o silicone oil o p e en he samples ~ - ----------------~,------- The e ec o addi ion o selec ed ca ageenans on iscoelas ic p ope ies o model S3 om dehyd a ion. Values o s o age modulus G', loss modulus G" and angen s o phase shi angle ( an 0 = G" /G') we e chosen o compa ison o cheese p o- pe ies. T ea men s we e s a is ically e alua ed by Wilcoxon es o compa ison o mean alues. Non-pa ame ical p ocedu es we e chosen due o a low amoun o a aila- ble da a (Ag es i, 1984). To show he signi icance, di - e ences among compa isons had o achie e P < O.OS. RESULTS AND DISCUSSION A i s , he heological analysis o model p o- cessed cheese om G oup I con aining 40% w/w DM and 4S% w/w o SO% w/w FDM was pe o med wi h he aim o e alua e he e ec o a dosing p ocedu e o 0.2S% w/w KC (in he o m o powde and aqueous suspension) on iscoelas ic p ope ies o he inal 10' .. e:. .. = -g ~ 10' b l:! " ...l "C = " C 10' :. .. .. .9 '" 10' 0.1 .. e:. :; "C ~ 10' b ~ ...l "C = " b 10' :. E ;l 10' 0.1 p oduc . The basic analysis e ealed ha all samples had p ac ically he same con en o DM (i.e. anging om 40.62 o 41.1S% w/w; P:::: O.OS) and ha also hei pH was e y simila (i.e. anging om S.86 o S.92; P:::: O.OS). These simila cha ac e is ics enabled o compa e he e ec o KC applica ion because bo h showed a high e ec on he consis ency o p ocessed cheese (Ma chesseau e a ., 1997; Lee e a ., 2004). As shown in Tab. I and Fig. 1 he o m, in which he K-ca ageenan is applied (i.e. ei he as a powde o as dispe sed in wa e p io o he applica ion in o he aw ma e ial), did no show any signi ican e ec on s o- age (G') and loss (G") moduli (P:::: O.OS). The e i- ciency o KC applied in concen a ion o O.2S% w/w was no dependen on i s physical o m and o ha eason i was decided o use i as a powde . F om he economic poin o iew his me hod o applica ion is easie and hus also cheape . A 10 B 10 F equency [Hz) 1: Dependence o s o age G' ( ull symbols) a loss G" (open symbols) moduli on equency o con ol sample (110), KC in powde o m (4D 0) a KC in aqueous dispe sion ( .... .6.) in g oup I o p ocessed cheeses wi h 45% w/w (A) and 50% w/w (B) FDM 54 M Ce nikowi, F. Buiika, V. Pa l nek, P. B ezina, J. H abe, S. K acma I: Values o s o age (G') and loss (G '') moduli and an 8 o e e ence equency 1 Hz in p ocessed cheese g oup I wi h addi ion o 0.25% w/w K-ca ageenan applied in powde o m and dispe sed in wa e * The a in d y ma e Fo m o applied G' G" an 8 con en o p ocessed cheeses (% w/w) K -ca ageenan [Pal [Pal [- ] none 596 ± 89.2 a 760 ± 54.6' 1.275 45 powde o m 1682 ± 167.7b 1397 ± 132.7b 0.831 dispe sed in wa e 1525 ± 159.1 b 1296 ± 116.3 b 0.850 none 145 ± 21.6' 284 ± 19.1 a 1.959 50 powde o m 612 ± 66.3 b 653 ± 53.5 b 1.067 dispe sed in wa e 522 ± 58.2b 590 ± 37.4 b 1.130 * S o age G' and loss G" moduli a e p esen ed by mean ± S.D.; an 8 = G" /G'. Means (n = 4) wi hin a column ollowed by no common supe sc ip le e di e (P < 0.05); samples wi h 45% w/w and 50% w/w FDM we e e alua ed sepa a ely. The ea e , he heological analysis o model p o- cessed cheese samples om G oups II o V wi h 40% w/w DM and 45% w/w o 50% w/w FDM was pe - o med. Bo h KC and IC we e applied in he a ge concen a ion o 0.25% w/w in he inal p oduc . The esul s o a basic analysis indica ed ha samples om all ou g oups unde s udy showed a p ac ically iden- ical con en o DM ( anging om 40.20-41.13% w/ w; P ~ 0.05) and also pH alues ( anging om 5.85- 5.93; P ~ 0.05). As shown in Tabs II and III and Figs 2 and 3, in which esul s o a dynamic oscilla ion heome y o G oups II-V wi h he con en s o 45% w/w and 50% w/w FDM a e p esen ed, he applied concen a ion o ca ageenans (0.25% w/w) inc eased signi ican ly he alues o bo h s o age (G') and loss (G") moduli (P < 0.05) wi hin he whole ange o es ed equencies (0.1-50Hz) as compa ed wi h co esponding con - ols. This e ec was obse ed in all es ed G oups (11- V) i espec i e o deg ee o ma u i y. These changes indica e ha he p esence o 0.25% w/w o bo h KC and IC in p ocessed cheese can change he cha ac- e o i s gel: as compa ed wi h con ol, he i mness o es samples was highe . This can be explained by he ac ha in e ac ions o bo h KC and IC chains con ibu e o he o ma ion o a mo e complex ma ix in samples o p ocessed cheese (Langendo e a!., 1999; Langendo e a!., 2000). II: Values o s o age (G') and loss (G '') moduli and an 8 o e e ence equency 1 Hz in p ocessed cheese g oups II and III wi h addi ion o 0.25% w/w K-ca ageenan (KC) and -ca ageenan (IC) applied in powde o m * The a in d y Type o G oup o p ocessed cheese ma e con en o added II III p ocessed cheeses ca ageenan G' G" an 8 G' G" an 8 (%w/w) [Pal [Pal [- ] [Pal [Pal [- ] none 2952 ± 153.5' 2267 ± 82.3' 0.767 1627 ± 76.6' 1436 ± 57.3' 0.882 45 KC 5835 ± 121.9b 3270 ± 127.1 b 0.561 3826 ± 112.9 b 2424 ± 287.4 b 0.635 IC 8714 ± 404.8 c 4445 ± 295.7c 0.510 3197 ± 156.2 c 2107 ± 268.5 b 0.659 none 1275 ± 102.2' 1329 ± 97.5 a 1.043 352 ± 32.3' 563 ± 62.4' 1.600 50 KC 4098 ± 99.7 b 2719 ± 132.0 b 0.664 641 ± 45.2b 721 ± 104.1 b 1.126 IC 5288 ± 275.3 c 2969 ± 186.4c 0.561 2750 ± 154.6c 1722 ± 243.4 c 0.627 * S o age G' and loss G" moduli a e p esen ed by mean ± S.D.; an 8 = G"/G'. Means (n = 4) wi hin a column ollowed by no common supe sc ip le e di e (P < 0.05); samples wi h 45% w/w and 50% w/w FDM we e e alua ed sepa a ely. The e ec o addi ion o selec ed ca ageenans on iscoelas ic p ope ies o model 55 III: Values o s o age (G') and loss (G '') moduli and an 8 o e e ence equency 1 Hz in p ocessed cheese g oups IV and Vwi h addi ion o 0.25% w/w K-ca ageenan (KC) and -ca ageenan (IC) applied in powde o m * The a in d y Type o G oup o p ocessed cheese ma e con en o added IV V p ocessed cheeses ca ageenan G' G" an 8 G' G" an 8 (%w/w) [Pal [pa] [- ] [pa] [Pal [- ] none 1509± 87.4' 1483 ± 91.8' 0.983 352 ± 10.1" 525 ± 16.8' 1.488 45 KC 2616 ± 48.6b 1958 ± 142.7b 0.748 1179 ± 132.4 b 1155 ± 23.5 b 0.979 IC 3450 ± 122.9 c 2387 ± 159.0c 0.692 1452 ± 60.0 c 1143 ± 16.1 b 0.787 none 178 ± 28.9' 337 ± 26.6' 1.889 46 ± 3.6' 128 ± 11.2' 2.762 50 KC 322 ± 76.0b 500 ± 37.1 b 1.552 597 ± 14.3 b 552 ± 18.7b 0.926 IC 1335 ± 129.5 c 1133 ± 82.8c 0.848 924 ± 69.2c 796 ± 25.6 c 0.863 * S o age G' and loss G" moduli a e p esen ed by mean ± S.D.; an 8 = G" /G'. Means (n = 4) wi hin a column ollowed by no common supe sc ip le e di e (P < 0.05); samples wi h 45% w/w and 50% w/w FDM we e e alua ed sepa a ely. 'i" e:. 10' 10' :; ~ b ~ 10' 10' 1 ~ , I! ~ A B 10' 10' 0. 10 0.1 10 ~ 10' 10' iI ~ ~ ! 1 10' 10' .. ~ k c 10' 10' 0.1 10 0.1 10 F equency [Hz) F equency [Hz) 2: Dependence o s o age G' ( ull symbols) a loss G" (open symbols) moduli on equency o con ol sample (110), KC in powde o m (eo) a lC in powde o m (A..L:,) o p ocessed cheeses wi h 45% w/w FDM (A -g oup 11, B -g oup Ill, C - g oup IV and D -g oup V). 56 M Ce niko a, F. Buiika, V Pa linek, P. B ezina, J. H abe, S. K acma 'ii' 10' 10' E!!. := .. ~ 6 10' 10' :a oS "0 Ii b 10' 10' ~ !! ~ A B 10' o· 10' 0.1 10 0.1 10 ~ 10' 'i ~ b 10' 10' ~ ...:I '8 II b 10' 10' i c 10' o· 0.1 10 0.1 10 F equency [Hz) F equency [Hz) 3: Dependence o s o age G' ( ull symbols) a loss G" (open symbols) moduli on equency o con ol sample (II 0), KC in powde o m (CD 0) a IC in powde o m (A. 1:::,.) o p ocessed cheeses wi h 50% w/w FDM (A -g oup IL B -g oup IlL C - g oup IV and D -g oup V). Tabs II and III and Figs 2 and 3 also indica e ha applica ion o IC in concen a ion o 0.25 % w/w esul ed in he o ma ion o a i me gel han ha o KC (P < 0.05). An explana ion o a be e e iciency o IC is no simple, especially wi h ega d o he indings pub- lished by !meson (2000) ha KC o med i m gels and IC so elas ic gels. The answe can be looked o e.g. in he ionic en i onmen (especially in he p esence o ions), which shows a signi ican e ec on he s eng h o ca ageenan ne wo k (Nicke son e al., 2004). In na u al cheese, which is he basic aw ma e ial o p o- duc ion o p ocessed cheese, he con en o calcium ions is app oxima ely en- imes highe han ha o po assium ions (Fox e aI., 2000). I is known ha IC is mo e sensi i e o po assium while KC is mo e suscep- ible o calcium ions (!meson, 2000). As men ioned by Spagnuolo e al. (2005) no only absolu e amoun s o calcium and po assium ions bu also he a io be ween hese ions may be impo an . Hence, dominance in con- cen a ion o Ca++ ions o e K+ ions may play a key ole in he i mness o KC and IC gels, espec i ely. In his con ex i is also app op ia e o ake in o accoun esul s published by MacA ain e al. (2003) who men- ion ha in case o KC he e is an op imal concen a ion o calcium ions in he medium and ha wi hin he ange o subop imal concen a ions hei inc easing con en esul s in a highe i mness o KC gel. Howe e , i he concen a ion o calcium ions su passed i s op imum wi hin a gi en sys em, any u he inc ease esul ed in a dec ease in alues o s o age (G') modulus (i.e. in he i mness) o he p oduced gel. CONCLUSION In his pape he e ec o addi ion o 0.25% w/w o K-ca ageenan and -ca ageenan on iscoelas ic p o- pe ies o model samples o p ocessed cheese con a- ining 45% w/w and 50% w/w a in d y ma e was e alua ed. I was ound ou ha a a a ge concen - a ion o 0.25% w/w o ca ageenans hei e ec on he consis ency o p ocessed cheese was no depen- den on he o m o applied addi i e ( es ed we e i s powde o m and i s aqueous dispe sion). The addi- ion o 0.25% w/w o bo h ca ageenan unde s udy esul ed in he o ma ion o i me and less sp eadable p oduc s in compa e o con ol samples wi hou ca - ageenan addi ion. The e ec o -ca ageenan was hi- ghe han ha o K-ca ageenan. The e ec o addi ion o selec ed ca ageenans on iscoelas ic p ope ies o model 57 SOUHRN Vli p ida ku yb anych ka agenanu na iskoelas icke las nos i modelo ych a enych sy u Vli pi'ida ku 0,25 % w/w K-ka agenanu a -ka agenanu na iskoelas icke las nos i a eny~~ ~y u byl sledo an u modelo y-ch zo kU s 40 % w/w susiny a 45 % w/w, esp. 50 % w/w uku susme. Ta e- ne sy y byly hodnoceny po 14 dnech sklado ani pi'i eplo e 6 ± 2°C. Sledo ane a ene sy y se nelisily zakladnich pa ame ech -susine a pH (P 2': 0,05). Nebyly zjis eny s a is icky y-znamne ozdily hod- no elas ickeho modulu p uZilos i G' [Pal, z a o eho modulu p uZilos i G" [Pal a angen u Uhlu azo- eho posunu an 8 [-] p o e e encni ek enci 1 Hz mezi a enymi sy y s aplikaci K-ka agenanu p as- ko e o me a e o me dispe ze e ode (P 2': 0,05). V dusledku pi'ida ku 0,25 % w/w K-ka agenanu i -ka agenanu ( p asko e o me) byl zaznamenan na us elas ickeho G' a z a o eho G" mod~lu p uz- nos i a pokles hodno an 8 (P < 0,05). Ta ene sy y se s aly uzsimi e s o nani s kon olniml zo ky bez pi'ida ku ka agenanu. Io a-ka agenan pi'idany p asko e o me byl p o koncen aci 0,25 % w/w pi'i z yso clni uhos i sledo anych a enych sy u liCinnejsi nez K-ka agenan (P < 0,05). a eny sy- , uk susine, kappa-ka agenan, io a-ka agenan, eologie This wo k was kindly suppo ed by a p ojec o Czech Minis y o Educa ion, You h and Spo s (G an No. MSM 7088352101). REFERENCES AGRESTI, A.: Analysis o o dinal ca ego ical da a. John Wiley & Sons, Inc., 1984,287 p. AUGUSTIN, M. A., PUVANENTHIRAN, A. and MCKINNON, 1. R.: The e ec o K-ca ageenan con o ma ion on i s in e ac ion wi h casein micelles. In . Dai y J., 1999,9: 413--414. ISSN 0958-6946. BERGER, w., KLOSTERMEYER, H., MERKE- NlCH, K. and UHLMANN, G.: P ocessed Cheese Manu ac u e. A JORA Guide. BK Giulini Chemie GmbH & Co. OHG Ladenbu g, 2002, 238 p. BOURRlOT, S., GARNIER, C. and DOUBLIER, J. L.: Micella -casein-K-ca ageenan mix u es. 1. Phase sepa a ion and ul as uc u e. Ca bohyd. 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