NG DNG PECTINASE LM TRONG RU VANG V NC QU

* Ru vang l sn phm ln men khng chng ct t nho. Ngy nay khi nim v ru vang c m rng ra cho cc loi qu nh m, mn, da, to, du,... Ru vang l loi nc ung c gi tr b dng cha lng etanol va phi (10 - 140GL) Ru vang, v mt l thuyt c chia lm 2 giai on ln men chnh: - Giai on 1 : Nh vo tc dng ca nm men Saccharomyces cerevisiae (S.cerevisiae) chuyn ho ng trong qu to thnh etanol v CO2 l ch yu. Ngoi ra, giai on ln men ny cn to ra mt s sn phm ph khc (axit hu c, ru cao phn t, este,...) - Giai on 2: (Cn gi l giai on ln men ph hay l giai on ln men malolactic). Nh vo tc dng ca vi khun lactic trong Leuconostoc oenos (L.oenos) c coi l tc nhn chnh yu ca giai on ny. Sn phm chuyn ho quan trng trong giai on ny l: Axit malic chuyn thnh axit lactic lm cho v ca ru vang tr nn chua du v "m". Axit xitric v ng qu (glucoza, fructoza) s to ra cc hp phn diaxetyl, axetoin v 2,3 batylen - glycol l t hp hng thm cho ru vang. Nm men khng chuyn ho axit hu c theo hng nu trn. * Nc qu (nc qu trong v nc qu c) l 1 trong nhng sn phm ch bin t qu thng qua vic tch dch qu bng nhiu phng php khc nhau. Vic lm trong dch qu trong thi gian ngn l rt kh khn bng phng php lng lc. Mc trong ca nc qu c vai tr n gi tr cm gic ca sn phm. 2.1. Mc ch: Pactimase (hay cn gi l pectinesterase) c tc dng hu hiu trong vic lm trong dch qu hoc sn xut nc qu trong hoc lm trong dch qu trc khi ln men sn xut ru vang. S dng pectinase s a hiu qu cao bi: Lng pectinase s dng t nhng hiu qu lm trong cao v nhanh thay th cho phng php ho - l (lng lc t nhin) km hiu qu, d nhim khun v thi gian di. 2.2. Nguyn tc:

Di tc dng ca pectinase, pectin - mt polysacharid c to thnh t s lin kt gia cc mch ca phn t axit galacturonic (C6H10O7) m mt phn ( mc khc nhau) c este ho vi ru metylic (CH3OH) b phn hu hoc b phn ct lin kt glucozit. Pectin l 1 cht in ly cao phn t (polime) mang in tch m (-) dc theo chiu di ca mch phn t c s sp xp cc nhm din tch cng du (COO-) nn mch c xu hng dui thng to nn b mt hp ph ln trong ton b dung dch c cha pectin. Nc qu c l h huyn ph trong phn t sn (tht qu) c kch thc nh, b mt ring ln v c nng lng rt ln nn khng bn vng v mt nhit ng hc. Mt khc, chng ta l nhng phn t tch in nn di tc dng ca lc ht phn t chng lin kt li to thnh nhng phn t ln d dng sa lng. Trong nc qu, s c mt ca hp cht polyphenol (c bit l tanin) to nn vi pectin nhng kt ta khng tan v.v... S c mt ca pectinase s thc y nhanh qu trnh c bn nu trn. Xc nh gi tr mt quang OD (Optical Density) = 600 nm theo thi gian so snh tc sa lng bt tht qu gia cc mu. 2.3. Phng php tin hnh: T nguyn liu qu ti (i, to, cam, qut,...) to ra nc qu dng c theo s tng qut sau: Nguyn liu Nc nng X l (ngm, ra) Trch ly b p Dch qu B Nng nhit Pha ch lc tinh Phn phi hnh tam gic y np

 un si, lm ngui Kim tra axit v Bx ca nc qu B sung pectinase pectinase 1% tinh khit Xc nh OD - Cc thng s cng ngh khi thao tc lm nc qu: + Ngm st trng bng dung dch clorin (CaOCl2) 5 mg/l trong 510 pht tu thuc vo mc nhim bn. + Dng my xay hoc my p tri cy lm nh nguyn liu. + Lc s b bng 4 lp vi mng hoc ry. + Nng nhit: nhit < 70 0C (t 65-68 0C) trong 2 - 3 pht nhm kt ta protein, cht cht ca qu. + Pha ch: Dng hn hp ng (hoc sir ng 70%), axit xitric, nc si ngui. Vi hn hp trn pha ch nc qu c Bx : 16-17 %; axit 0,25 - 0,3 % (tnh theo axit xitric). + Lc tinh: qua 24 lp vi mng. + Phn phi hnh tam gic: dng phu sch phn phi mi bnh 100 150 ml, y kn bng nt nhm hoc bng khng thm nc (6 bnh/1 nhm). + un si; lm ngui: Dng ni cch thu un si dung dch v gi nhit ny trong thi gian 25-30 pht, lm ngui nhanh n nhit mi trng trong nc tan. + B sung pectinase 1 % vi liu lng 3 % vo 2 bnh; 5 % vo 2 bnh, cn li 2 bnh khng b sung pectinase (cc bnh tam gic cha nc qu). * Chun b mu thc nghim: 6 Mu nc qu c chun b sn trong bnh tam gic c dung tch 100 - 150ml sn phm. Dng pectinase ca hng Novo (an Mch) 1%, c cho vo: + 3% enzim pectinase (theo th tch nc qu) : 2 bnh + 5% enzim pectinase (theo th tch nc qu : 2 bnh + 0% enzim pectinase (mu i chng) : 2 bnh Cc bnh c lc u trong cng 1 thi gian, chia u lm 2 loi mu: 1 loi yn theo di trong sut qu trnh thc nghim; 1 loi ly mu o OD theo thi gian (gi): 0, 1, 3, 5, 7 bc sng = 600 nm. * Tnh kt qu v biu din kt qu:

 bc sng = 600 nm, tin hnh o gi tr OD cc mu i chng v thc nghim. Ly gi tr OD o c gi sau so vi gi tr OD o c thi gian k trc , chng ta c gi tr OD. OD l gi tr biu t bin dao ng ca tr s OD gia cc ln o theo thi gian. OD cng nh chng t s bn vng ca h huyn ph nc qu c cng cao. Ngc li, gi tr OD tnh c cng ln, tc sa lng bt tht qu cng nhanh chng t s tch pha rn cng nhanh trong h nc qu c. iu ny ng ngha vi vai tr lm trong ca pectinase. Kt qu OD c biu din trn h trc to hoc bng profil bin theo thi gian gia cc mu thc nghim (k c mu i chng). So snh v nhn xt. 2.4. Nguyn liu, dng c v ho cht: - Nguyn liu: + Qu ti (i, to, cam, qut,...) + Axit xitric tinh th thc phm + ng knh trng (RS hoc RE) + Axit ascocbic (vitamin C) trng (nu dng to, l, i) + Pectinase ca hng Novo (an Mch) - Dng c: + My p tri cy hiu philippe hoc National + Ry lc + Khn lc loi 4 lp v 24 lp (mi nhm 1 khn mi loi). + Phu nh + Bnh tam gic loi 200ml (6 ci/1 nhm) c nt y chit quang k cm tay. + My so mu c bc sng = 600 nm v cc cuvet. + Chai l sch c nt nhm cha enzim pectinase. + Bng sch (thm nc v khong thm nc) + Ni cch thu (mi nhm 1 ni) + Nhit k 1000C (mi nhm 1 chic) + Buret, pipet, bnh tam gic 50ml. - Ho cht: + Clorin (CaOCl2) + NaOH 0,1 N + Phenolphtalein