The Influence of Glumes on Malting and Brewing of Hulled Wheats

The Influence of Glumes on Mlting nd Brewing of Hulled Whets Ombrett Mrconi, 1 Deprtment of Economic nd Food Science, University of Perugi, Vi Sn Costnzo, 06126, Perugi, Itly; Heidi Myer, Itlin Brewing Reserch Centre, University of Perugi, 06126, Perugi, Itly; Frncesco Chicchieroni nd Eros Ricci, Deprtment of Economic nd Food Science, University of Perugi; nd Giuseppe Perretti nd Polo Fntozzi, Itlin Brewing Reserch Centre, University of Perugi, 06126, Perugi, Itly ABSTRACT J. Am. Soc. Brew. Chem. 71(1):41-48, 2013 The composition nd mlting behviors of different mlts nd worts of the hulled whet vrieties Rossorubino (Triticum dicoccum) nd Monlis (T. monococcum) mlted with nd without glumes were studied. During mlting, crospires were removed together with the rootlets fter kilning from the dehulled smples, wheres only the rootlets were removed from the hulled smples. The mlt nlysis reveled tht the hulled mlts exhibit lower soluble protein content with higher free mino nitrogen nd totl free mino cid vlues. The mount of the dimethyl sulfide precursors ws lower in the dehulled mlts thn in the hulled ones. Bsed on its positive mlt prmeters, Rossorubino ws chosen to produce three beers: 100% hulled Emmer mlt beer nd, mixed with brley mlt, 40% hulled Emmer mlt beer nd 40% dehulled Emmer mlt beer. The technologicl prmeters of the reltive beers were in the stndrd rnge. The 100% hulled Emmer mlt beer showed lower polyphenol content thn the brley mlt beers nd hd good fom stbility. The descriptive sensory nlysis showed tht ll descriptors were present in the 100% Rossorubino beer with medium-low scores. All pnelists observed refreshing chrcter with notes of honey nd florl, citric rom of the beer. Keywords: Brewing, Glumes, Lignns, Mlting, Triticum dicoccum, Triticum monococcum. RESUMEN El comportmiento de mltedo y composición de ls mlts diferentes y mostos de l vriedd de trigo cscdo Rossorubino (Triticum dicoccum) y Monlis (Triticum monococcum) mlt con y sin glums fueron estudidos. Durnte el mltedo, ls plúmuls fueron retirron junto con ls ricills después del secdo de ls muestrs descscrdos, mientrs que sólo ls ricills se hn retirdo de ls muestrs con cscrill. El nálisis de mlt reveló que ls mlts con cscrill exhiben menor contenido de proteín soluble con myor FAN y los vlores totles de minoácidos libres. El importe de los precursores de dimetil sulfuro (DMS-P) fue menor en ls mlts descscrdos que en los con cscrill. Bsándose en sus prámetros de mlt positivos, Rossorubino fue elegido pr producir tres cervezs: un cervez de 100% de mlt Emmer cscdo, y mezcldo con mlt de cebd, un cervez de 40% de mlt Emmer cscdo, y un cervez de 40% de mlt Emmer descscdo. Los prámetros tecnológicos de ls reltivs cervezs estbn en el rngo norml. L cervez de 100% de mlt Emmer cscdo mostrron un menor contenido de polifenoles de l cervez de mlt de cebd y tení buen estbilidd de l espum. El nálisis sensoril descriptivo mostró que todos los descriptores estbn presentes en el cervez de 100% Rossorubino con medio-bjos puntjes. Todos los pnelists observó un crácter refrescnte con nots de miel y un rom florl, cítrico de l cervez. Plbrs clves: Elborción de l cervez, Glums, Lignnos, Mltedo, Triticum dicoccum, Triticum monococcum 1 Corresponding uthor. Phone: +39 075 585 7926; Fx: +39 075 585 7946; E-mil: omrconi@unipg.it http://dx.doi.org/10.1094/asbcj-2013-0124-01 2013 Americn Society of Brewing Chemists, Inc. Hulled whets such s Triticum dicoccum (emmer) nd T. monococcum (einkorn) re mong the most ncient of cerels. They belong to the fmily Pocee, genus Triticum, nd re cultivted whet species like the more common T. estivum nd T. durum. Einkorn whet hs diploid number of chromosomes (2n = 14) while emmer whet is tetrploid (2n = 28) (28). Hulled whets re chrcterized by tough, lrge glumes which give them dded protection in the field nd during storge. The cultivtion of hulled whets originted in the Middle Est (Irn, Irq, Turkey, Syri, nd Plestine) bout 10,000 yers go nd spred during the Neolithic period into Europe. During the Romn period, emmer whet ws mjor food source but, tody, hulled whets re underutilized crops becuse they hve lrgely been replced by free-threshing whets which re esier nd cheper to process nd provide higher yields (17,44). In modern griculture, emmer nd einkorn whet re mostly found in orgnic griculture (22). They hve lso been noted to possess positive nutritionl nd helth-promoting properties such s improving glycemic control nd lowering cholesterol levels, s well s hving high ntioxidnt, vitmin, nd minerl contents (4,16,36,37). Unsurprisingly, these whets re becoming incresingly sought fter by consumers. Tody, einkorn whet is grown in Western Turkey, the Blkn countries, Itly, Spin, Switzerlnd, nd Germny (42). Emmer whet remins n importnt crop in Ethiopi nd minor crop in Indi, Itly, nd Turkey (30). In Itly, these whets hve been commonly consumed in regionl specilties such s slds nd soups but more nd more emmer nd einkorn whets re being used in the production of bred, biscuits, nd pst, nd emmer whet is now sold in nerly every Itlin supermrket (8,9). Recently, locl breweries in Germny nd Itly hve ttempted to crete new beer styles with these ncient whets in the interest of developing new nd different products nd to ensure tht the consumer hs lrge selection in flvors nd tstes of beer. At the sme time, vrious scientific studies were conducted tht confirmed the suitbility of emmer nd einkorn whets s rw mterils for the brewing nd mlting industry. In these studies, different mlting procedures were used, For exmple, Zrnkow et l (45) nd Jentsch (20) mlted the cerel without the husk while Myer et l (32) mlted with the husk. In generl, the mount of husk is between 20 nd 30% of the whole weight of the cerel (32). Normlly, prior to the processing of grins for soups, slds, nd flour, the glumes re eliminted. The procedure is simple mechnicl dehulling process, lthough it does require investments of time nd money. Dehulling is generlly recommended for mlting nd brewing s well (33) becuse, especilly when husk content is high, the yield is smller. This hs been confirmed by the nlysis of the extrct of the Congress wort (32). Furthermore, the presence of the husk could chnge the flvor nd technologicl prmeters of the wort. In fct, ccording to some uthors, thick husks of brley mlt contin more polyphenols nd bitter substnces which cn negtively ffect colloidl stbility nd tste of the finl beer (18,26). Moreover, in contrst to hulled brley, which is used commonly in breweries nd hs husks tht 41

42 / Mrconi, O., Myer, H., Chicchieroni, F., Ricci, E., Perretti, G., nd Fntozzi, P. strictly dhere to the cryopsis, the dehulling of emmer nd einkorn whets is comprtively esy nd cn be done with suitble equipment nd cre to void the mechnicl or microbiologicl dmge of the germ (17). However, the hull cn be useful during mlting for protecting nd conserving the crospires nd lso during lutering, when the hulls form cke tht cts s filter (15). Lignns re polyphenols found in plnts tht exhibit vrying estrogenic, ntiestrogenic, nd ntioxidnt ctivities (2). With the help of the intestinl microflor, they cn be converted in the intestine into enterolignns which my protect ginst certin types of cncer nd crdiovsculr disese (34). Lignns re found primrily in flxseeds, nuts, cerels, legumes, berries, nd fruits (7,40). The only dt bout the lignn content of T. dicoccum, s totl secoisolriciresinol, hs been reported s 8.1 µg per 100 g (1), while there is no informtion in the literture bout the content of lignns in mlts nd beers mde from hulled whet. Other reserch on lignns in rye nd whet indicte tht they my influence the flvor of these cerels (10,13,19,27). An objective evlution of the contribution of the hull to wort composition cn only be mde by mlting nd brewing emmer nd einkorn whets with nd without the hull nd compring the results. The purpose of this study ws to determine the different mlting behviors nd compositions of mlts from the Rossorubino vriety of emmer whet (T. dicoccum) nd the Monlis vriety of einkorn whet (T. monococcum), mlted with nd without glumes. We lso investigted the brewing behvior of nontrditionl cerels by compring 100% emmer mlt beer (brewed with hulled mlt), brley mlt beer with 40% hulled emmer mlt, nd brley mlt beer with 40% dehulled emmer mlt. Becuse mlting releses bioctive compounds due to the formtion nd ctivtion of enzymes (13,23,27) the lignn content ws lso ssessed, s were other prmeters, including totl nd soluble nitrogen content, free mino nitrogen (FAN), Kolbch index, nd pprent limit of ttenution. EXPERIMENTAL Apprtus An utomtic micromlting system from Custom Lbortory Products (Scotlnd) ws used s previously described (32). Unmlted Cerel Smples The unmlted grin smples were supplied by Prometeo srl (Urbino, Itly), nd cultivted in centrl Itly. The T. dicoccum vriety Rossorubino ws hrvested in 2009 nd the T. monococcum vriety Monlis ws hrvested in 2008. The dehulling process ws performed by the producer. The smples were stored t 4 C before their use. The brley mlt ws obtined from commercil supplier nd the mlting qulity ws confirmed by nlyticl determintion. Grin Qulity Prmeters Qulity prmeters in the grins were determined using the Anlytic-EBC methods (12) for the following vribles: grin moisture (%), EBC method 3.2; protein content (% d.m.), EBC method 3.3.1 (totl N 5.7 for whet nd totl N 6.25 for brley); nd germintive energy nd wter sensitivity, EBC method 3.6.2. Wter Uptke by the Hulled nd Unhulled Cerels Wter uptke during steeping of the hulled mlt ws studied in previous rticle (32). For the dehulled mlts, the two smples were steeped in 15 C wter for 18 hr. Every hour, wter uptke ws determined by mesuring the weight difference of the smples. This nlysis ws performed in duplicte. Mlting Procedure The mlting procedures were lso performed in duplicte. The hulled grins were mlted following the procedure described in previous work (32) using the following prmeters: steeping-out moisture, 47%; steeping wter temperture nd temperture of the ir during rest, 15 C; first steeping durtion, 4 hr; ir rest, 20 hr; second steeping durtion, 2 hr. For the dehulled grins, the following prmeters were used: steeping wter temperture, 15 C; first steep durtion, 1 hr; then, the steeping wter ws chnged; second steep durtion, 7 hr. A steeping-out moisture of 40% ws chieved. For ll trils for both hulled nd dehulled grins, the vegettive period ws 120 hr (steeping nd germintion) nd the kilning regime ws 15 h, t 55 C, 4.5 h, t 72 C, nd 3.5 h, t 82 C. After drying, the rootlets nd crospires were removed from the dehulled mlts wheres only the rootlets were removed from the hulled mlts becuse the crospires were protected from the glumes. Mlt Anlysis Mlt qulity ws ssessed by nlysis of the mlt in duplicte with the following Anlytic-EBC (12): moisture content (%), EBC method 4.2; extrct yield (% d.m.) nd scchrifiction time (min), EBC method 4.5.1; totl N content (% d.m.), EBC method 4.3.1; soluble N content (% d.m.), EBC method 4.9.1; Kolbch index; FAN content (mg/100 g), EBC method 4.10; viscosity (mps t 20 C nd 8.6 P), EBC method 4.8; pprent finl ttenution (%), EBC method 4.11.1; disttic power (WK), EBC method 4.12; ph, EBC method 8.17; mlt color (EBC-U), EBC method 4.7.1; nd totl polyphenols (Congress wort), EBC method 9.11. Free mino cid content in the crude grins, mlt, Congress worts, nd beers. Asprtic cid, glutmic cid, serine, threonine, rginine, histmine, methionine, vline, leucine, isoleucine, lysine, glycine, lnine, tyrosine, nd phenyllnine were determined in the crude grins, mlt, Congress worts, nd beers by HPLC quntifying the fluorescence of the orthophtldildehyde (OPA)/mecptoethnol derivtives (29). Aliquots (1 g of cerel or 1 ml of wort or beer) were extrcted with 10 ml of 5% (w/v) trichlorocetic solution for 30 min with stirring. Then, the smple ws centrifuged t 3,000 rpm for 5 min nd the superntnt ws pssed through 0.45-µm filter, derivtized with OPA in the presence of mercptoethnol, nd nlyzed by HPLC coupled with fluorimetric detection (Agilent, Snt Clr, CA). The detector ws set t λ emission of 330 nm nd λ excittion of 420 nm. The chromtogrphic seprtion ws performed by C 18 Inertsil ODS 3 (Vrin) (150 by 4.6 mm, 5 µm) using the following grdient: t = 0 min 100% A, t = 30 min 70% A, t = 43 min 48% A, t = 45 min 30% A, nd t = 60 min 0% A. Solvent A ws 0.05 M solution of K 2 HPO 4 (81%), methnol (18%), nd tetrhydrofurn (THF) (1%) t ph 7.5 nd solvent B ws 0.05 M solution of K 2 HPO 4 (30%) nd methnol (70%). The flow rte ws set t 1 ml/min nd the loop ws 20 µl. The mino cids were identified by comprison of their retention times with those of the stndrds mix for fluorescence detection (Fluk, Buchs, Switzerlnd) nd they were quntified by using their clibrtion curves in the liner rnge of 0.781 12.500 nmol/ml. Clibrtion curves were determined for six different concentrtions of stndrd solutions. Ech clibrtion smple ws injected in triplicte nd clibrtion grphs for ech compound were obtined for the different concentrtions by plotting concentrtion ginst pek re. The coefficient of determintion ws r 2 > 0.990 nd the repetbility (RSD%) of the methods ws lower thn 14% for ll the mino cids. Lignns in Congress worts nd beers. The secoisolriciresinol diglucoside (SDG), isolriciresinol (ISOLARI), hydroxymtiresinol (HYDROXY), secoisolriciresinol (SECO), lriciresinol

Glumes in Mlting nd Brewing / 43 (LARI), pinoresinol (PINO), nd mtiresinol (MATA) in wort nd beer were extrcted nd purified following solid-phse extrction method (3) nd nlyzed by HPLC coupled with coulometric detection (14). Two Jsco PU-1580 pumps were used, connected to grdient solvent system, Bsic Mrthon Sprk (Erkerode, The Netherlnds) utosmpler with 100-µL loop, n Inertsil ODS-3V column (C 18 250 by 4.6 mm Ø; prticle size 5 µm), CoulArry detector (ESA Inc., Chelmsford, MA), nd softwre for Windows for cquisition nd elbortion of dt. The eight electrode potentils were 100 905 mv t increments of 115 mv. Mobile phse A ws 0.05 M KH 2 PO 4 nd 0.05 µm sodium luryl sulfte (SLS) nd mobile phse B ws phse A/CH 3 - OH/CH 3 CN, 30:20:50 (v/v/v), 0.05 µm SLS. The mobile phses were djusted to ph 3.35 with 85% orthophosphoric cid nd were filtered with 0.22-µm membrne filter (Type GSWP for queous solvents, GVWP for orgnic solvents; Millipore, Bedford, MA). The voltmmetric dt were collected nd nlyzed by CoulArry softwre. The lignns were identified by comprison of their retention times with those of stndrds nd they were quntified by using their clibrtion curves in the liner rnge of 50 1,050 µg/l. The coefficient of determintion (r 2 level) ws bove 0.99 nd the repetbility (RSD%) of the method ws lower thn 3% for ll the nlytes. Totl dimethyl sulfide nd dimethyl sulfide precursor contents. The totl dimethyl sulfide (DMS) nd dimethyl sulfide precursor (DMS-P) contents were determined ccording to the method from Stfisso et l 2011 (41). Brewing The mlt smples were processed in 20-L pilot-scle brewery (Brumeister, Speidel, Germny) in duplicte. Mlt (3.5 kg) ws crushed in two-roller mill (120 kg hr 1 cpcity; Engl, Engl- Mschinen GmbH, Schwebheim, Germny) with gp of 0.5 mm between the rollers. The grist ws then mixed with 14 L of brewing wter t 52 C. Mshing ws executed by incresing the temperture by 1 C /min nd mintining the following rests: 30 min t 52 C, 45 min t 65 C, 20 min t 72 C, nd 20 min t 76 C. Afterwrd, the msh ws filtered in luter tun. Sprging ws executed twice with 3 L nd once with 2 L of brewing wter t 78 C. Next, the msh ws boiled for 90 min until reching pproximtely 12 P. At the beginning of this phse, hop pellets (Styrin Golding vriety, 46 g/kg [w/w] of α-cids) were dded to chieve 20 Interntionl Bitter Units (IBU). After boiling nd elimintion of the hot trub, the wort ws cooled to 15 C. Aertion of the wort ws chieved. Fermenttion Pitching of the yest ws crried out t 15 C using 11.5 g of commercil English le yest (Sfle Fermentis, S-04, Mrcq-en- Broeul Cedex, Frnce) fter rehydrtion with sterile wter. The pitching rte ws 3.9 10 6 cells/ml. The fermenttion temperture ws mintined t 16 C for 3 dys. After cooling, when the finl ttenution ws chieved, the mturtion lsted for week t 4 C. At the sme time, the dischrge of the yest from the bottom of the tnk ws performed. After this phse of mturtion, the beer ws refermented in the bottle with the ddition of sucrose (4.5 g/l) nd plced in thermostt t 20 C for 1 week. At the end of the re-fermenttion, the beer ws stored t 4 5 C until nlysis. Wort Anlysis The following nlyses were performed in duplicte ccording to the officil Anlytic-EBC methods (12): extrct ( P), EBC method 8.3; ph, EBC method 8.17; color (EBC-U), EBC method 8.5; viscosity (mps t 20 C), EBC method 8.4; fermentbility, ttenution limit (%), EBC method 8.6.2; totl nitrogen (mg/l), EBC method 8.9.1; nd FAN (mg/l), EBC method 8.10. Beer Anlysis The beer smples were chrcterized for the following prmeters in duplicte by the Anlytic-EBC methods (12): originl, rel, nd pprent extrct nd originl grvity ( P), EBC methods 9.4 nd 9.43.2; lcohol (% v/v), EBC method 9.2.1; ph, EBC method 9.35; color (EBC-U), EBC method 9.6; totl Nitrogen (mg/l), EBC method 9.9.1; FAN (mg/l), EBC method 9.10; fom stbility (NIBEM 30 s), EBC method 9.42; nd totl polyphenols (mg/l), EBC method 9.11. Sensory Descriptive Anlysis Sensory tests were crried out with trined, 12-member tsting pnel (2 women nd 10 men ged 25 50 yers old). The pnelists were trined in the identifiction nd determintion of the globl intensity nd primry descriptors of beer (mlt chrcter, hop chrcter, ester chrcter, sweetness, bitterness, body, nd ftertste). A score of 0 ment tht the descriptor ws bsent wheres score of 5 indicted tht the descriptor ws extremely strong. Some comments bout secondry descriptors were lso expressed by the pnelists. Sttisticl Anlysis Sttisticl nlysis ws performed using SigmStt softwre (version 11; Jndel Scientific, Sn Rfel, CA). Comprisons of the different mtrices were mde by one-wy repeted mesures nlysis of vrince, nd the results were further nlyzed using the Holm-Sidk test. The unpired t test ws used t P < 0.05 to determine sttisticlly significnt difference between two mtrices. RESULTS AND DISCUSSION In the current study, two hulled whet species, T. dicoccum nd T. monococcum, were mlted with nd without the glumes. Mlting the grin with the hull offers the dvntge tht the crospires nd the germ re protected during mlting but the cerels re wter sensitive. This mens tht the germintive energy is gretly decresed when more wter is vilble thn is needed for germi- TABLE I Grin Qulity Prmeters of Triticum dicoccum (Rossorubino) nd T. monococcum (Monlis) Monlis Rossorubino Hulled Dehulled Hulled Dehulled Moisture (%) 9.3 12.4 b 12.0 c 12.6 d Germintive energy (%) 100 92 99 92 Wter sensitivity (%) 51 89 b 87 b 92 b Totl protein (N 5.7) (% d.m. b ) 10.5 13.9 b 11.3 c 12.3 d Vlues in the sme row followed by different letters re sttisticlly different (P < 0.05). Number of repetitions: 2. b Dry mtter.

44 / Mrconi, O., Myer, H., Chicchieroni, F., Ricci, E., Perretti, G., nd Fntozzi, P. ntion. Generlly, the wter sensitivity depends on the vriety, growing re, climtic conditions, nd husk (33). In this study, hulled nd dehulled cerels from the sme btch were used, so the wter sensitivity depended only on the presence of glumes (Tble I). We found tht, during mlting of the hulled smples, n ir rest of 20 hr between the two steeping times ws necessry to overcome this issue. In this cse, the turning of the grin hppened utomticlly. In contrst, the dehulled whet is not wter sensitive (Tble I). This behvior is opposite to the wter sensitivity reported for whet (33). As result, the first steep of 1 hr ws relly only clening process. The second steep lsted 7 hr, until the steeping-out moisture of 40% ws chieved. No ir rest ws done. The smples were turned by hnd no more thn two times per dy to preserve the crospires nd germs. The dehulled grins showed decresed germintive energy (Tble I), probbly due to dehulling, which ws performed by the supplier. The totl protein content ws 10.5 13.9%. These vlues re cceptble, even if the vlue for the dehulled Monlis smple (13.9%) ws slightly incresed from norml. Generlly, mximum protein content of 11.5% for brley nd 12.5% for whet re required for brewing (33). The mlting qulity prmeters of the T. monococcum nd T. dicoccum hulled nd dehulled mlts re reported in Tble II. The fine extrcts of the dehulled Monlis nd Rossorubino mlts were clerly higher thn their respective hulled mlts becuse the glumes contributed minor mount to the extrct, negtively influencing the yield. The dehulled Monlis nd Rossorubino mlt extrct vlues re similr to the extrct vlue of stndrd whet mlt, which generlly hs no glumes (>83%) (20,24,33). The color of ll the mlts ws between 5.3 nd 5.7 EBC units, excluding the hulled Rossorubino mlt, which ws higher (7.4 EBC). Rossorubino mens ruby red, nd it seems tht these coloring substnces re locted in the glumes. The wort viscosities of the hulled nd dehulled Monlis mlts were very high in comprison with brley or whet mlt while the wort viscosities of hulled nd dehulled Rossorubino mlt were comprble with brley mlts. These dt re in greement with dt of previous study (20). The dehulled mlts of ech vriety hd higher viscosity vlues thn the hulled mlts. The pprent finl ttenution ws higher in the hulled mlts, with the vlue for Rossorubino similr to those of brley mlts nd even better thn reported, perhps due to the different vrieties used (20). Monlis, on the other hnd, hd very low vlues of pprent finl ttenution nd this could be correlted with its low disttic power vlues. The totl protein contents, rnging from 9.8 to 12.8%, were considered cceptble for brewing (11,31,38). All mlts hve high content of soluble nitrogen which is greter in dehulled mlts thn hulled mlts, indicting high proteolytic ctivity. For whet mlt, excessive proteolytic ctivity could be negtive for the sensoril qulity, resulting in low rom (6,25). For ll the smples, the trend of the soluble nitrogen vlues ws higher thn the stndrd commercil requirements for brley nd whet mlts nd greed TABLE II Qulity Prmeters for Triticum dicoccum (Rossorubino) nd T. monococcum (Monlis) Mlts Monlis Rossorubino Prmeters b Hulled Dehulled Hulled Dehulled Brley Whet (32) Fine extrct (% d.m.) 66.4 86.4 b 68.1 c 88.3 d 81.1 e >83 ph 6.09 6.04 6.05 5.97 5.90 b Color (EBC-U) 5.5 5.7 7.4 b 5.3 3.6 c 4.0 Scchrifiction time (min) 15 10 <10 15 <10 <10 Viscosity 8.6 P (mps) 2.06 2.96 b 1.54 c 1.64 c 1.51 c <1.8 Apprent finl ttenution (%) 73.4 65.8 b 83.9 c 78.6 d 79.4 e 81 Totl protein (N 5.7) (% d.m.) 9.8 12.8 b 10.1 c 12.3 d 11.5 e (N 6.25) 11.5 12.5 Kolbch index 40 49 b 41 63 c 37 38 Soluble nitrogen (mg/l) 8.6 P 883 1,094 b 910 1,312 c 736 d 700 750 FAN 8.6 P (mg/l) 158 107 b 163 131,b 144,b 120 130 Disttic power 211 234 287 b 264 c 294 b >250 Totl polyphenols 8.6 P (mg/l) 36 22 b 54 c 40 66 c Totl free mino cids 8.6 P (mg/l) 678 467 b 780 c 655 Totl lignns 8.6 P (mg/l) 4.14 4.64 b 1.80 c 2.08 d Totl DMS (mg/kg d.m.) 23.3 14.9,b 21.6 6.7 b DMS-P (mg/kg d.m.) 9.4,b 9.2,b 15.8 4.5 b Vlues in the sme row followed by different letters re sttisticlly different (P < 0.05). Number of repetitions: 2. b Abbrevitions: d.m. = dry mtter; FAN = free mino nitrogen; DMS = dimethyl sulfide. TABLE III Comprison of the Amino Acid Profile (mg/kg d.m.) of the Crude Cerels nd Their Corresponding Mlts Cerel, mlt Asp Glu Ser His Arg Gly Thr Al Tyr Met Vl Phe Leu Ile Lys Totl Rossorubino Hulled 149 269 34 24,e 74 34 34,c 92 32 68 35 14,c 13 10 33,c 917 Hulled mlt 199 b 377 b 146 b 167 b 441 b 85 b 188 b 284 b 193 b 116 b 240 b 259 b 164 b 163 b 138 b 3,160 b Dehulled 187 b 310,b 43 25,e 74 40 33,c 92 32 82 38 11 16 12 23 1,019 c Dehulled mlt 26 c 269 60 52 c 63 44 46 193 c 44 37 c 84 c 60 c 55 c 69 39 c 1,143d Monlis Hulled 188 b 200 c 33 21 70 38 25 c 61 d 32 44 c 29 10 18 9 25,c 804 e Hulled mlt 312 d 425 d 264 c 179 d 699 c 104 c 234 d 322 e 247 c 157 d 308 d 346 d 276 d 345 c 230 d 4,448 f Dehulled 252 e 297,b 40 31 e 98 d 34 28,c 61 d 36 78 64 c 30,c 49 c 23 97 e 1,218 d Dehulled mlt 85 f 333 b 189 d 115 f 268 e 94 b,c 145 e 327 e 132 d 109 b 205 e 197 e 159 b 188 b 103 e 2,664 g Brley mlt 359 452d 198 d 80 g 178 f 13d 108 f 221 f 282 e 38 c 405 f 415 f 209 e 342 d 164 f 3,464 h Vlues in the sme column followed by different letters re sttisticlly different (P < 0.05). Number of repetitions: 4. Abbrevitions: d.m. = dry mtter; Asp = sprtic cid; Glu = glutmic cid; Ser = serine; Thr = threonine; Arg = rginine; His = histmine; Met = methionine; Vl = vline; Leu = leucine; Ile = isoleucine; Lys = lysine; Gly = glycine; Al = lnine; Tyr = tyrosine; Phe = Phenyllnine.

Glumes in Mlting nd Brewing / 45 with the totl protein vlues (39), such tht high contents of totl protein corresponded to high vlues of soluble nitrogen. Conversely, the FAN vlues showed the opposite trend: the hulled mlts, with less soluble nitrogen, presented clerly higher FAN vlues thn the dehulled ones. To further confirm the result of the FAN vlues, the totl free mino cid contents of the cerels nd the respective mlts were evluted. The totl free mino cid content of the hulled cerels ws very low in comprison with the dehulled ones but incresed with mlting from 917 to 3,160 mg/kg for Rossorubino nd 804 to 4,448 mg/kg for Monlis (Tble III). The dehulled crude cerels hd higher mino cid contents thn the hulled ones, but did not increse very much during mlting: 1,019 to 1,143 mg/kg for Rossorubino nd 1,218 to 2,664 mg/kg for Monlis. This behvior of the mino cid nd FAN vlues probbly ws connected to the loss of the crospires together with the rootlets of the dehulled mlts when clened fter kilning wheres, for the hulled mlts, only the rootlets were eliminted. It is likely tht significnt mount of free mino cids nd FAN is locted in the ctive tissues of the crospires. Asprtic cid nd glutmic cid re the min mino cids present in the cerels. During mlting, the mount of sprtic cid in the dehulled mlts decresed clerly with respect to the other mino cids. The rginine content of Rossorubino nd, especilly, of Monlis hulled mlt ws much higher thn in brley mlt. On the other hnd, the vline, isoleucine, nd leucine contents of ll the whet mlts were lower thn in brley mlt. It is known from the literture tht low content of these mino cids is n indictor of incresed dicetyl production during fermenttion (35). The DMS-P of the emmer nd einkorn mlts were significntly higher thn expected for brley mlt (<5 mg/kg) (26), with the exception of Rossorubino dehulled mlt. This could be due to the high mount of methionine in these smples. It is well known tht one of the DMS precursors, S-methyl-methionine (SMM), is synthesized by L-methionine S-methyltrnsferse (SMM synthetse) during germintion simultneously in the germ nd rootlets from S-denosyl-L-methionine nd methionine (5,43). The dehulled smples without crospires nd rootlets hd lower vlues compred with the hulled ones. The mount of totl polyphenols in Congress worts of the hulled mlts were surprisingly low compred with the brley mlt wort despite the gret content of the hulls. This trend ws confirmed in the 100% Rossorubino mlt wort. The lignn contents in the Congress worts were 4.14 nd 4.64 mg/l for Monlis nd 1.80 nd 2.08 mg/l for Rossorubino in the hulled nd dehulled grins, respectively. In both species, the lignn content of the dehulled mlt ws higher (Tble II). Mshing The Rossorubino mlt ws chosen to produce top-fermented beer becuse its technologicl prmeters re in the rnge of suitble brewing mlts in comprison with brley nd whet mlt requirements. The Monlis vriety ws less suitble to mlting due to its high mlt viscosity, high protein modifiction, pprent limit ttenution, nd disttic power. As is well known, high viscosity with high protein modifiction do not permit for compenstion for the mlt qulity with n dequte mlting technology (6). For the 100% Rossorubino beer, the mlt ws milled with two-roller mill. The glumes remined nerly intct, wheres the grin ws well ground. This might ccount for the low content of polyphenols in the finl beer (Tble IV) becuse the less the husk is broken, the less substnces like polyphenols would be extrcted. During mshing, the glumes bsorbed lrge mount of wter, with n optiml wter/grist rtio of t lest 4:1. The volume of the spent grins per kilogrm of milled cerel ws quite high: 2.73 L/kg for emmer mlt compred with 1.56 L/kg for brley mlt. As result, emmer mlt needed lot of sprging wter to reduce the extrct of the second wort. Alterntively, the residul extrct of the lst run ws high nd the corresponding yield ws low. The lutering ws excellent, with filtrtion rte of 1.8 L/min compred with 0.6 L/min in the brley mlt wort. The glumes of the hulled whet were very good filter mteril for both the 100 nd 40% hulled Rossorubino mlt beers, with filtrtion rtes of 1.8 L/min. This is becuse the husk cretes less compct cke thn pure brley mlt, llowing for fster filtrtion. The opposite hppened with the dehulled Rossorubino mlt, such tht dding 40% dehulled mlt to the grist cused lower filtrtion rte becuse of less spent grins nd higher viscosity. In this cse, the filtrtion rte ws 0.4 L/min. The nlyticl prmeters of the resulting worts re shown in Tble IV. The soluble protein content of the wort ws high for the 100% Rossorubino hulled mlt, 40% hulled Rossorubino mlt, nd 40% dehulled Rossorubino mlt worts t 1,530, 1,227, nd TABLE IV Technologicl nd Qulity Prmeters of the Worts nd Beers Obtined with Triticum dicoccum (Rossorubino) Mlt Prmeters 100% Hulled mlt 40% Hulled mlt 40% Dehulled mlt 100% Brley mlt Wort Extrct ( P) 11.5 12.5 b 12.0 c 13.0 d ph 5.50 5.50 5.08 b 5.19 b Color (EBC-U) 14.9 7.1 b 7.4 b 7.6 b Viscosity (mps) 1.84 1.79 1.78 1.79 Apprent fermentbility (%) 86.0 88.0 84.4 85.3 Totl nitrogen (mg/l, 12 P) 1,530,c 1,227 b,c 1,381 c 1,145 b FAN b (mg/l, 12 P) 206 179 b 197 c 221 d Beer Originl extrct ( P) 11.4 12.9 b 12.3 c 12.5 c Apprent extrct ( P) 2.1 1. 9 b 1.8 c 2.2 d Rel extrct ( P) 3.9 3.8 3.9 4.1 b Alcohol (% v/v) 4.90 5.43 b 5.43 b 5.16 b Color (EBC- U) 10.8 8.0 b 6.4 c 7.6 b ph 3.98 4.23 b 4.23 b 4.17 b Totl nitrogen (mg/l, 12 P) 903 799 b 879 823 b FAN (mg/l, 12 P) 45 36 b 37,b 158 c Fom stbility (Nibem 30 sec) 252 191 b 146 c 171 b,c Totl polyphenols (mg/l) 99 137 b 137 b 210 c Vlues in the sme row followed by different letters re sttisticlly different (P < 0.05). Number of repetitions: 2. b Free mino nitrogen.

46 / Mrconi, O., Myer, H., Chicchieroni, F., Ricci, E., Perretti, G., nd Fntozzi, P. 1,381 mg/l, respectively, wheres it ws 1,145 mg/l for the 100% brley mlt wort. The FAN content ws quite low, between 179 nd 206 mg/l for the Rossorubino mlts compred with 221 mg/l for the brley mlt; nevertheless, the fermenttion ws not influenced negtively. The fermenttion ws completed fter four dys. The pprent fermentbility ws good for ll the smples. Brewing The technologicl prmeters of the reltive beers were in the stndrd rnge. The color decresed from the 100% Rossorubino hulled mlt beer to the 40% Rossorubino hulled mlt beer nd, finlly, to the 40% dehulled Rossorubino mlt beer. A lower polyphenol concentrtion for the Rossorubino beers compred with TABLE V Amino Acid Profiles of Wort nd Beer Obtined with 100% Triticum dicoccum (Rossorubino) Mlt Profiles (mg/l) Amino cids b Wort Beer Group A Asp 56 1 b Glu 61 1 b Ser 66 n.d. Thr 49 6 b Arg 169 3 b Group B His 52 1 b Met 20 0.1 b Vl 97 0.2 b Leu 62 0.4 b Ile 120 n.d. Lys 93 3 b Group C Gly 43 1 b Al 102 8 b Tyr 73 24 b Phe 116 0.2 b Totl 1,180 49 b Vlues in the sme row followed by different letters re sttisticlly different (P < 0.001); n.d. = not determined. Number of repetitions: 4. b Abbrevitions: Asp = sprtic cid; Glu = glutmmic cid; Ser = serine; Thr = threonine; Arg = rginine; His = histmine; Met = methionine; Vl = vline; Leu = leucine; Ile = isoleucine; Lys = lysine; Gly = glycine; Al = lnine; Tyr = tyrosine; Phe = Phenyllnine. the 100% brley mlt beer is shown in Tble IV, confirming the lower polyphenol content for emmer whet in the Congress wort. The mino cid profiles of the 100% Rossorubino hulled mlt wort nd beer divided into groups (A, B, nd C) with respect to the mino cid uptke by yest during fermenttion, s suggested by Jones nd Pierce (21), is shown in Tble V. Almost ll the mino cids were consumed by the yest t the end of the fermenttion. The totl lignn content of the 100% Rossorubino mlt beer ws 2.31 mg/l in the wort nd decresed in the finl beer to 0.561 mg/l (Tble IV). There ws 76% loss of the lignns during brewing. No dt re present in the literture bout the lignn content of beer. This study found tht the lignn content in the 100% Rossorubino mlt wort ws mostly secoisolriciresinol diglucoside nd isolriciresinol. Hydroxymtiresinol, secoisolriciresinol, lriciresinol, nd pinoresinol were present in low mounts while mtiresinol ws bsent (Tble VI). In beer, the lignns were represented minly by secoisolriciresinol diglucoside nd secoisolriciresinol. At the end of the mturtion time, the 100% Rossorubino hulled mlt beer nd the 100% brley mlt beer were subjected to pnel test with 12 trined tsters who evluted the sensory profile of the beers (Fig. 1). The descriptive sensory nlysis TABLE VI Lignn Concentrtion in Wort nd Beer Obtined with 100% Triticum dicoccum (Rossorubino) Mlt Concentrtion (µg/l) Lignn b Wort Beer SDG 1,179 417 b ISOLARI 883 n.d. HYDROXY 29 n.d. SECO 80 98 LARI 33 15 PINO 99 32 b MATA n.d. n.d. Totl 2,310 561 b Vlues in the sme row followed by different letters re sttisticlly different (P < 0.001); n.d. = not determined. Number of repetitions: 4. b Abbrevitions: SDG = secoisolriciresinol diglucoside, ISOLARI = isolriciresinol, HYDROXY = hydroxymtiresinol, SECO = secoisolriciresinol, LARI = lriciresinol, PINO = pinoresinol, MATA = mtiresinol. Fig. 1. Spider web plots of scores (0 5) on primry descriptors of 100% emmer mlt beer.

Glumes in Mlting nd Brewing / 47 showed tht ll studied descriptors were present in the 100% Rossorubino beer, with scores in the medium to low rnge. All pnelists observed refreshing chrcter with notes of honey nd florl, citric rom to the beer. A slight senstion of stringency ws perceived. The beer presented the typicl turbidity of whet beer nd hd good fom stbility (250 Nibem t 30 sec). Generlly, ll the pnelists greed tht the body of the beer ws wek. The 40% Rossorubino hulled mlt beer nd 40% Rossorubino dehulled mlt beer were lso subjected to screening, which gve positive evlution for these beers. In conclusion, we found tht both T. dicoccum (vr. Rossorubino) nd T. monococcum (vr. Monlis) cn be successfully mlted with nd without the glumes. The min difference between the two mlting procedures ws the elimintion of the crospires together with the rootlets fter kilning in the cse of the dehulled smples wheres, in the cse of the hulled smples, only the rootlets were eliminted becuse the crospires were covered by the glumes. This leds to higher FAN, pprent limited fermentbility, nd free mino cid contents in the hulled smples. The reltively high content of methionine could cuse high content of DMS-P. Bsed on the good mlt prmeters, the vriety Rossorubino ws chosen to produce finished beers. The mlt without the glumes cnnot be used becuse it lcks hulls for the wort filtrtion. The high mount of glumes in the hulled smple did not led to high polyphenol content in the wort or beer. The hulled emmer mlt cn be used to produce 100% hulled emmer mlt beer tht is refreshing nd pltble. The Monlis vriety of einkorn whet ws not suitble for brewing 100% Einkorn beer. 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