NG DNG CA ENZYMEE PECTINASE Trong sn xut thc phm, ngi ta thng s dng cc ch phm pectinase di dng tinh khit.

Ngi ta khng dng ch phm di dng canh trng nm mc sy kh. T lng ch phm pectinase c c trn lng nguyn liu em ch bin vo khong t 0.03 0.05 n 0.10%. Pectinase thng c s dng trong cc ngnh cng nghip thc phm sau: Sn xut ru vang; Sn xut nc qu v nc ung khng c cn. Sn xut cc mt hng t qu: nc qu c c, mt nh, mt ng, Sn xut nc gii kht; Sn xut c ph v c ph ha tan.

Trong sn xut ru vang, cng nh trong sn xut nc qu v cc nc ung khng ru, u c th s dng pectinase mt cch rt hiu qu. Nh tc dng ca pectinase m cc qu trnh p, lm trong v lc dch qu rt d dng, do lm tng hiu sut sn phm. Chng hn a pectinase vo khu nghin qu, s lm tng hiu sut nc qu sau khi p ln ti 15 25%. Bi l khi c pectin th khi qu nghin s c trng thi keo, do khi p dch qu khng thot ra c. Nh pectinase phn gii cc cht pectin di m dch qu trong sut khng b vn c v lc rt d dng. Khng nhng vy, enzyme pectinase cn gp phn chit rt c cc cht mu, tanin v nhng cht ha tan na, do lm tng thm cht lng ca thnh phm. Trong sn xut cc mt hng t qu (mt nh, mt ng, ) pectinase cng c vai tr quan trng. Nh pectinase m c th thu c dch qu c nng m c. Chng hn nh dch to c c n 72 Brix, nu khng tch cc pectin t nhin cha trong i th sn phm s b keo t mt cch mnh m v khng th c c hn na. a s trng hp, ngi ta kh pectin i, sau mi lc ri c c, nhng i khi ngi ta cho pectinase tc dng trong sut thi gian c c.

Trong sn xut c ph, ngi ta dng pectinase tch lp keo trn b mt ca ht c ph. Trc y ngi ta dng ngay vi sinh vt lm cng vic ny, nhng thng qu trnh xy ra khng ng u v kh kim tra. Hin nay ngi ta thng dng cc ch phm pectinase. Vic ng dng pectinase sn xut nc qu c p dng ln u tin vo nm 1930. T n nay vic p dng enzyme ny tr nn rt ph bin nhiu nc trn th gii. Vic sn xut nc qu t trc n nay ch yu bng phng php p. Nu pectin cn nhiu s theo nc qu v gy ra hin tng nc qu b c, c keo cao v rt kh lc trong. Trong t bo ca qu nc chim khong 90 95%. Nu ta ch nghin sau p th ta ch c th sn xut c khong 60 70% l ti a. Khi ta cho enzyme pectinase vo, hiu sut p s tng 15 30%. Nhiu trng hp, hiu sut p tng n 50%. Liu lng ch phm enzyme tinh khit cho vo l 0,03 0,05% hoc ch phm th l 0,5 2%. Nhit duy tr cho qu trnh thy phn l 43 45oC. Thi gian thy phn l 4 8h. dch qu thu c bng pectinase s trong hn, kh nng lc s tt hn v hiu qu kinh t thy r. C CH TC NG CA ENZYME PECTINASE Nguyn tc lm trong dch qu: 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. T cc loi qu khc nhau, ngi ta thng ch bin thnh cc dng nc qu trong, nc qu c v phm, trc tin phi chit rt c dch qu t m c bn, m b

i khi m c hc na. Do , hiu sut chit rt dch qu s ph thuc vo kh nng thm ca t bo, cu to gii phu v tnh cht c l ca nguyn liu, nht ca chnh dch qu, chc ca tht qu, cng nh thnh phn nh tnh v nh lng ca pectin cha trong dch bo v trong thnh t bo ca m qu. s dng vo vic ch bin nc qu trong th ch phm pectinase phi c enzym endo v exo-polygalacturonse, enzym pectinesterase v proteinaza. Trong ch phm loi ny th nhng enzym va k trn l nhng enzym quyt nh hiu qu tc dng ca ch phm. V endo v exo-PG s lm gim nht ca dch qu; cn enzym PE cng gp phn vo tc dng ca enzym ny. Enzym proteinase ca ch phm s thy phn v protein ca v nguyn sinh t bo, kt qu l lm cho s thot ca dch qu d dng. S c mt ca xenluase v hemixenlulase trong ch phm cng tt nhng khng bt buc. Khi s dng cho cc loi qu nh to, l th trong ch phm khng c php c enzym pectinaseliminase, v enzym ny s phn hy protopectin vn gn cc t bo ring bit ca m qu li vi nhau, do gy mn ho m qu, lm tng nht ca dch qu v kt qu l lm gim hiu sut dch qu. Ngc li, khi s dng cho cc loi qu mng nh du ty th s c mt trong ch phm enzym PTE li cn thit v enzym ny s lm mn ho t bo lm cho tnh cht thot nc ca t bo tng ln cn nht ca dch qu li gim xung. Ngoi ra, i vi cc ch phm pectinase dng trong sn xut nc qu trong cng khng c php c cc enzym oxydase (ascohatoxydase, poliphenoloxydase). Trong ch bin nc qu c tht th cc enzym PTE, hemixenlulase v xenlulase li l nhng enzym quyt nh hiu qu tc dng ca ch phm. Trong vai tr dn ng l enzym PTE c tc dng phn hy protopectin ca m qu. Hemixenlulase v xenlulase s cng vi PTE lm cho ng th ca nc qu c tht tt hn. Trong trng hp ny s c mt trong ch phm enzym PG c bit l endo-PG l khng cho php v l ng th ca nc qu s ph thuc vo nghin mn ca m qu v nht ca dch qu. Trong ch phm loi ny c mt proteinase cng tt nhng khng bt buc. Ngoi ra khi dng cc nguyn liu c mu sng th trong ch phm khng c

cha enzym oxi ho, cn cho cc nguyn liu c carotenoid v antoxian th khng c cha enzym ph hy cc cht ny. thu c nc qu, trong trng hp c s dng enzym pectinase, ngi ta cng phi nghin nh nguyn liu qu. B nghin nhn c em x l bng enzym, sau mi em p hoc ly tm. Trong trng hp c s dng enzym, ngi ta c th nghin nh ti mc ti a, ct sao thun li cho tc dng ca enzym m khpng s b tc nghn knh dn dch qu trong qu trnh p sau ny. V l sau khi kinh qua tc dng ca enzym, dch qu d dng thot ra khi b nghin, nn khi p hoc ly tm, ch lm cng vic tch dch khi cc tiu phn ca m qu m thi. Thc t sn xut chng t khi x l b to nghin bng 0,03% ch phm pectinase hiu sut ca dch qu c nng ln t 20-25%. Hoc khi thu nc nho m khng s dng ch phm pectinase th hiu sut dch ch t c ti a l 65% so vi khi lng nguyn liu. Nu s dng pectinase th tng hiu sut dch qu t 65% ln 77,3% (nho trng) v t 66% ln 82,2% (nho ). Dng pectinase khng ch lm tng hiu sut dch qu m cn lm trong dch qu. Phng php s dng enzym c hiu qu hn c. Bi l nh tc dng ca cc enzym m cc h keo ca nc qu s b ph hu hon ton. Nu nh khi lm trong bng cc phng php khc pectin b kt ta mt phn th khi x l bng pectinase, pectin b phn gii hon ton thnh cc cht ho tan. Vi phng php enzym, khi mang mt lng tha ch phm s loi tr c trng hp lm trong khng hon ton. Dch nc qu c x l bng ch phm pectinase thng c v hon ton hn v t c khuynh hng b c hn. Qu trnh lm trong dch qu di tc dng ca pectinase c th chia lm ba giai on:

Giai on bt n nh ho c tiu biu bng s gim nht mt cch su sc v thng c gi l trng thi ph v. T bo thc vt c cu to bng v t bo (thnh t bo). V t bo nh mt lp thnh bo v rt hu hiu v to hnh cho t bo. v t bo thc vt c nhiu cht pectin, cc cht pectin c xem nh cht ciment gn cc t bo vi nhau. Ph v s gn kt ny s to iu kin cho cc vt cht trong t bo thot ra khi t bo. Cc ch phm enzyme c cha khng ch pectinase m cn cha cc enzyme trong nhm cellulase. Cc loi enzyme ny s lm ph v thnh t bo v gip qu trnh sn xut dch t bo tt hn. Giai on kt lng bt u t trng thi ph v v kt thc khi kt ta hon ton. Nc qu sau khi c tch khi t bo thng cha nhiu cht khc nhau. Trong

cht pectin chim lng ng k v pectin thng gy hin tng nht cao v gy c nc qu. Cc cht protein c trong bo tng, mng t bo v gian bo. Pectin cha polygalacturonic acid, araban v galactan. Trong lng polygalacturonic acid chim ti 40-60%. Khi b thy phn, pectin tch thnh hai phn: Phn trung tnh phc cht galactanoraban Phn acid acid pectic.

Trong bo tng, pectin nm dng ha tan. Trong mng t bo v gian bo, chng nm dng khng ha tan gi l protopectin. Protopectin mng gian bo c cha lng kim loi kh ln v mt lng nhm metocyl lm protopectin bn vng. Cn protopectin mng t bo cha mt lng kim loi khng nhiu, c metocyl ha cao. V th, t bo thc vt c kh nng trng n tt. Nu enzyme tham gia phn gii pectin gian bo s lm cc t bo kh lin kt vi nhau v tht qu d dng b mm ra. Pectin thng c mi lin kt hydro v lin kt nguyn t yu hn so vi cellulose. Tham gia phn hy pectin gm nhiu loi enzyme.

Giai on cui cng thng c xc nh bng s vng mt cc pectin kt ta bi ion canxi.

Pectin l hp cht polyme ca axit galaturonic, ester methyl v mt s thnh phn nh khc ch tn ti trong t bo thc vt. Carbohydrat tn ti cng pectin bao gm: araban, arabinoxylan, gaclactan. Pectin chia ra lm hai loi: loi tan trong nc v loi khng tan trong nc. Loi khng tan trong nc cn c gi l protopectin v chng thng lin kt vi cellulose to thnh pectinocellulose. Tn ti 3 nhm enzyme pectinase phn gii pectin:

Pectin methyl esterase Depolymease Exoenzyme pectinase

Pectin methyl esterase (EC.3.1.1.11): Enzyme ny tham gia phn gii pectin thnh axit pectic v methanol. Enzyme ny cn c tn khc l pectiestenase (PE). Hiu sut thy phn pectin ca enzyme ny rt cao, c th t c 98%. Pectinesterase ca nm si hot ng mnh pH 3 5. C hai loi pectinase ny b bin tnh nhit 80OC. Enzyme pectin methyl esterase tham gia tc ng vo COOCH3 theo c ch sau: Pectin depolymease: Enzyme ny c trong t bo thc vt. Tuy nhin hin nay ngi ta sn xut enzyme ny theo qui m cng nghip t nm si v t vi khun. Chng c phn ra lm 4 loi nh sau: 1-Endo pectin traseliminase (EC.4.2.2.3). vit tt l PTE pectiliase. 2-Endo pectin axit traseliminase (EC.4.2.2.1). vit tt l PATE. 3-Endo - poly galacturonase (EC.3.2.1.15). vit tt PG. 4-Endo - poly methyl galacturonase. vit tt PMG. Cc enzyme thng phn ct lin kt glucozit trong cu to ca pectin:

Enzyme polygalacturonase tham gia tc ng vo glucoside cui cng to ra nhng phn t cu thnh. Ty theo ngun sn xut enzyme depolymease m chng c pH ti u khc nhau. Ta c th tham kho pH ti u ca cc enzyme ny trong bng sau: STT 1 2 3 4 5 6 7 Ngun enzymee Aspergillus niga Flavobacterium Pectinovorum Asperigillus niger Aspergillus sp. Coniothyrum diplodiela Aspergillus sojae Becterium polymyxa Loi enzymee Endo PMG Endo PMG Endo PG Endo PG Endo PG Endo PTE Endo PLE pH ti uu ca Enzymee depolymease 5- exoenzyme l nhng enzyme gii phng axit galacturonic bt u t im cui ca phn t pectin. Enzyme pectinase c ng dng ch yu ph v thnh t bo ca qu nhm gii phng cc cht c trong t bo ca qu ra ngoi. pH ti u 5,2 7,4 8,2 4,7 4,8 3,7 4,2 4,4 5,5 8,9 9,1

Hiu qu ca vic s dng enzym pectinase So snh b sung pectinase v khng b sung pectinase trong qu trnh sn xut nc qu p B sung pectinase Khng b sung pectinase

Hiu sut thu dch p cao hn (tng 14% - Hiu sut thp hn. 30%). nht ca dch qu gim mnh. Qu trnh lc tt v nhanh hn. Tng nhanh qu trnh lm trong dch qu. Hiu qu ca vic s dng enzym nht dch qu cao. Qu trnh lc km v lu hn. Qu trnh lm trong dch qu mt nhiu thi gian.

 c nhiu nghin cu v kt qu p dng trong sn xut nc qu v ru vang cho thy khi s dng enzym pectinase cho hiu sut nc qu v cht lng ru vang rt cao. Khi tin hnh ng dng enzym trong sn xut nc qu v ru vang, ngi ta c bit quan tm n hai yu t c tnh cht quyt nh n hiu qu enzym. Nguyn liu Kh nng xay, nghin v lm nh nguyn liu.

Nguyn liu. Khng phi tt c cc loi qu u cho hiu sut cao khi x l enzym. Khi s dng enzym trong ch bin nc qu v ru vang ngi ta phi a ra chn k thut cho ph hp. Khi nim ny hon ton khc vi khi nim chn k thut khi trong cng ngh ngi ta khng s dng enzym. chn k thut sn xut nc qu l giai on chn ca qu, m bo tch dch qu c tt vi s tch t ti a cc cht c gi tr dinh dng cao v hng v thch hp. Tuy nhin cng cn bit rng, phn ln nguyn liu, hiu sut dch qu cao khng trng hp vi s tch t cc cht c gi tr dinh dng. nhiu loi qu, kh nng thot dch qu khng trng vi mc chn. Nhiu khi qu tht chn, kh nng tch t cht dinh dng cao nhng li khng thun li cho vic tch nc qu. Chnh v th, tng loi qu, ngi ta phi tin hnh phn loi v xc nh chn k thut ring, sao cho ph hp vi vic s dng enzym v thu hi dch qu. Cng vic th hai sau khi phn loi qu, vic nht thit phi lm trc khi s dng enzym l qu phi c lm sch bng cch ra nhiu ln loi b nhng tp cht, VSV. Sau l cc qu trnh k thut nh x nh, nghin, tng kh nng tc ng ca enzym v tng hiu sut thu dch qu. Kh nng lm nh nguyn liu. Lm nh nguyn liu l khu k thut rt quan trng, n quyt nh n hiu qu tc ng ca enzym v hiu sut sn xut dch qu cng nh cht lng dch qu.

Ty theo tnh cht c l v c im cu to ca tng loi qu ngi ta chn nhng phng php x l qu thch hp nh nghin ct, ch hay x nh. Khi thc hin mt trong nhng phng php c hc trn, mt s t bo b ph hy, mt tnh cht bn thm, do vic chn phng php c hc lm nh qu ph hp vi tng loi qu rt c ngha cng ngh. Nhng bt li khi s dng enzym Nc qu c s dng enzym c nhiu u im, tuy nhin vic s dng enzym cng c nhng nhc im nh xy ra nhng bin i bt li v VSV. Nu x l trong thi gian ngn, qu trnh oxy ha v s nhim VSV khng thnh vn ln. Nhng nu x enzym trong thi gian di s xy ra qu trnh oxy ho v s nhim VSV. Ngi ta ngn nga qu trnh oxy ho trong nc qu bng mt trong ba cch sau: Cho ascorbic acid Cho anhydric sulfur Cho enzym glucose oxidase. Trc khi dng nhng cht chng oxy ha trn, ngi ta phi un nng dch qu. Khi un nng dch qu, cc enzym c trong nc qu s b c ch. Theo , nc qu c ly tm lm sch c hc, sau nc qu c un nng ti 80OC v sau h nhit n nhit ti u ca hat ng enzym. Ngi ta thng cho lng ascorbic acid ti 5mg/100g nc qu hn ch qu trnh oxy ho. hn ch s pht trin ca VSV trong tng hp s dng enzym x l nc qu, ngi ta thng p dng nhng bin php sau. S dng sorbic acid, ascorbic acid, benzoate natri. Sorbic acid thng c s dng trong nc qu vi liu lng l 0,03%. Acid ny hot ng pH rt rng (pH 3,99,2).Sorbic acid c ch s pht trin ca nm men v nm si, khng lm thay i v v

mi ca sn phm. Benzoate natri thng c s dng trong nc qu vi liu lng l 0,15%. S dng anhydric sulfur. Ngi ta thng s dng anhydric sunfur bo qun nc qu bn thnh phm. Cht ny va c kh nng tiu dit VSV, va c kh nng chng oxy ho. Khi cho cht anhydric sunfur vo dch qu, chng d dng b oxy ha. Khi lng oxy c trong dch qu t dn v n mc trit tiu, gy ra trng thi th hiu oxy ha-kh trong dch qu thay i v VSV s b c ch hoc tiu dit. Tuy nhin cn phi lu rng, nu lng anhydric sunfur qu nhiu s dn n c ch cc hot ng ca enzym. Cc th nghim cho thy, nu hm lng anhydric snfur 0,1% lm gim hot tnh ca ch phm enzym, nu 0,5% th enzym hon ton b c ch. X l nhit. Ngi ta c th un nng dch qu bo qun nc qu. Tuy nhin, vic un nng dch qu khng th qu cao v chnh nhit s lm thay i tnh cht cm quan, thnh phn ha hc v hot tnh enzym. a s enzym phn gii pectin b mt hot tnh 70OC v chng hot ng mnh 45-50OC. Tuy nhin, nc qu ln men, ngi ta s dng enzym x l nc qu ngay nhit thng (trng vi nhit ln men).