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Enzym

Amylase

I. nh ngha v cng thc cu to Amylase l mt h enzyme rt ph bin trong th gii sinh vt . Cc enzyme ny thuc nhm enzyme thy phn, xc tc phn gii lin kt ni phn t trong nhm polysaccharide vi s tham gia ca nc: RR + H-OH -> RH + R-OH C cht tc dng ca amylase l tinh bt v glycogen. 1. -amylase (-1,4-glucanohydrolase): (EC 3.2.1.1) Amylase c kh nng phn ct cc lin kt 1,4glucoside ca c cht mt cch ngu nhin v l enzyme ni bo. -amylase khng ch c kh nng phn hy h tinh bt m cn c kh nng phn hy cc ht tinh bt nguyn vn 2. -amylase (glucoamylase) (EC 3.2.1.3) Glucoamylase c kh nng thy phn lin kt -1,4 ln -1,6-glucoside, ngoi ra cn c kh nng thy phn lin kt -1,2 v -1,3-glucoside. Glucoamylase c kh nng thy phn hon ton tinh bt, glucogen, amylopectin, dextrin thnh glucose m khng cn c s tham gia ca cc loi enzyme amylase khc. 3. enzyme isoamylase(glycogen 6glucanohydolase)(EC 3.2.1.68)

4. Enzyme pullulanase(-dextrin6- glucosidase)(EC 3.2.1.41) Khc vi dextrinase ti hn trong hot ng ca n trn glycogen, v t l ca thy phn dextrins gii hn. Tc ng ca n trn Amylopectin hon tt.Maltose l ng nh nht m n c th chuyn ha mi lin kt.

5. enzyme isoamylase(glycogen 6- glucanohydolase

II.

Phn loi Enzym chia thnh 2 nhm: Endoamylase ( -Amylase)

. -1,4-glucano hydrolase . -1,6-glucano hydrolase Exoamylase . -amylase . Amyloglucosidase (glucoamylase)

III.

Nguyn l hot ng ca enzym Enzym Amylase l enzym tinh bt, ng vai tr quan trng trong cng nghip thc phm. C ch tng qut nh sau: -Amylase Dextrin +oligosaccharid Tinh bt Dextrin Pulluulanase -Amyolase Maltose Dextrin Glucose Amyloglucosidase Trong sn xut cn khi thy phn tinh bt, -amylase tc ng vo lin kt 1,4 glucoside v sn phm to ra mantose v dextrin m mch ca chng gn bng C6. Cc dextrin sau s thy phn ch yu theo: C6 C7 C8 C5 +C1 C1+ C6 C2+C6 hay C5 + C2 hay C3 + C5

-amylase ca vi khun thy phn tinh bt tao ra lng glucose va maltose theo t l 1:5,45. -amylase ca nm si thy phn tinh bt to ra lng glucose v maltose theo t l 1:3,79. -amylase ca vi khun v mm si hon ton khng c kh nng phn gii -1,6 glucoside ca c cht.trong dch ha, pH ca dung dch bt thng trng hp vi hot ng ti u ca -amylase, pH ny cng l pH hot ng ti u khi tin hnh ng ha. Chnh v th nu

trong trng hp s dng c cc loi bt c pH trng vi pH hot ng ti u ca -amylase th phi iu ha pH cho thch hp Vai tr ca glucoamylase trong sn xut cn: Glucoamylase thy phn lin kt -1,4 trong cc polysaccharide, chng lin kt ct cc gc glucose khng kh trong mch polysacchride. Ngoi ra glucoamylase c c kh nng ct lin kt -1,6 glucoside Glucoamylase thng hot ng trong mi trng acide (pH hot ng ti u l khong 3,5-5,5). Ngoi ra cc glucoamylase cn c th hot ng trong mi trng trung tnh (pH 6,0-7,5). Cc glucoamylase ca nm men thng l nhng glucoamylase trung tnh. Phn ln cc glucoamylase hot ng pH khng cao. Nhit ti u khong 55-600C. sn phm cui cng ca hot ng glucoamylase l glucose, glucose c xc nh l ng ln men. Do , vic s dng glucoamylase trong sn xut cn c trin vng nht Bng. Mt s tnh cht ca glucoamylase Khong pH ti u 3,0-6,0 Ti u 4,5-5,0 2 Aspergillus awamori 2,7-7,0 Ti u 3,5-5,0 3 Rhizopus niveus 3,0-6,0 Ti u 3,5-5,0 Nhit ti u 55-60 Mt hot tnh 90 0C 55-65 Mt hot tnh 85 0C `55-60 Mt hot tnh 70-80 0C

Stt 1

Ngun enzym Aspergillus niger

VI. Ngun thu nhn v phng php thu nhn enzym 1. Ngun thu ng vt: Enzyme amylase c trong ty tng ca ng vt. Thc vt: c ly t nhiu ngun nh: i mch (hodeum sativum):

+ ging 2 hng: sn xut malt lm bia + ging nhiu hng (4,6 hng): lm thc n cho gia sc. La (oryza sativa L.): ch yu vng ng Nam . Ng (zea mays): c nhiu loi ng: ng , ng bt, ng rng nga. Ht ng c mu trng, mu vng hay mu hng. Ng mu vng do c s hin din ca carotenoid v zeaxanthine trong ni nh. Ng mu hng l do trong ni nh ca ng c anthocyanin Vi sinh vt: (VSV) Ngy nay do u th v nhiu mt, vsv tr thnh ngun thu enzyme amylase ch o. Nhng chng vsv to nhiu amylase thng c phn lp t cc ngun t nhin. Vsv to amylase c dng nhiu hn c l nm si,gi nm men v vi khun,cn x khun th t hn. Cc ging nm si thng dng l ging nm si Aspergillus, rhizopus Nm men v gi nm men thuc cc ging Candida, Saccharomyces, Endomycopsy, Endomyces cng to amylase. Nhiu vi khun c kh nng to lng ln amylase nh: Bac.polymyxa, Phytomonas destructans, Cassavanum cc vi khun a nhit c kh nng sinh trng nhanh v pht trin tt nhit cao nn khi nui chng t b nhim vsv khc. Trong nhm x khun rt him gp loi to amylase mnh m, tuy nhin cng c mt s t nh x khun a nhit. Micromonospora vugaris 42 c kh nng to mt lng nh a-amylase hot ng 65C cng vi protease v cc enzyme khc.

2. Phng php thu nhn Enzyme amylase Amylase, mt loi enzym co ngha v mt sinh l, thng mi v lch s cn c gi l diastase. Enzym ny c c trong thc vt ln ng vt. Amylase c tinh sch t Malt vo nm 1835 bi anselme Payen v Jean Persoz. Amylase thuc nhm Enzym thy phn, xc tc t phn gii cc lin kt glucoside ni phn t trong cc polysaccharide vi s tham gia ca nc. Amylase thy phn tinh bt, glycogen v dextrin thnh glucose, maltose v dextrin hn ch. Amylase c trong nc bt cn c gi l ptyalin. Amylase phn b rng ri trong t nhin v l mt trong nhng loi enzym c ng dng rng ri trong cng nghip. y t v nhiu lnh vc khc, c bit trong cc ngnh cng nghip thc phm lm lng tinh bt. Amylase thu nhn t ht ny mm, nm mc, nm men, vi khun. Trong Amylase c thu nhn t malt vi s lng rt nht, ch yu dng trong sn sut bia Enzym Glucoamylase. Enzym thuc nhm exo- -1,4D.Glucan glucohydrolate. Theo l thuyt, enzym ny c thy phn tinh bt to thnh glucose c hiu sut n 100% Vi sinh vt: (VSV) Ngy nay do u th v nhiu mt, vsv tr thnh ngun thu enzyme amylase ch o. Nhng chng vsv to nhiu amylase thng c phn lp t cc ngun t nhin. Vsv to amylase c dng nhiu hn c l nm si,gi nm men v vi khun,cn x khun th t hn.

Cc ging nm si thng dng l ging nm si Aspergillus, rhizopus. Nm men v gi nm men thuc cc ging Candida, Saccharomyces, Endomycopsy, Endomyces cng to amylase. Nhiu vi khun c kh nng to lng ln amylase nh: Bac.polymyxa, Phytomonas destructans, Cassavanum cc vi khun a nhit c kh nng sinh trng nhanh v pht trin tt nhit cao nn khi nui chng t b nhim vsv khc. Trong nhm x khun rt him gp loi to amylase mnh m, tuy nhin cng c mt s t nh x khun a nhit. Micromonospora vugaris 42 c kh nng to mt lng nh a-amylase hot ng 65C cng vi protease v cc enzyme khc. Phng php thu nhn malt: Malt i mch:

i mch

Thu nhn, x l, lm sch, phn loi

Sy kh

Bo qun

ngm

Khng kh Malt kh bo qun Ny mm

Sy kh

Tch mm r

Mm r kh

Malt thc:

Ht thc, la

Lm sch, phn loi

Sy bo qun

Ngm ht

Tch mm

malt

mm

Sy malt

malt

Phng php tinh sch amylase: 2 phng php. + Tinh sch enzyme amylase bng phng php sc k. + Thu nhn enzyme amylase bng phng php in di trn gel polyacrylamide (polyacrylamide Gel Electrophoresis _ PAGE) Phng php thu nhn t VSV: S tng hp enzyme amylase khng nhng ph thuc vo tnh cht di truyn ca VSV m cn ph thuc vic tuyn chn cc iu kin nui c hiu, ngoi ra cc iu kin l ha trong qu trnh nui cng ng vai tr quan trng.

Nhng yu t nh hng :thnh phn mi trng,tnh cht c l ca mi trng, tit trng, m ban u, thong kh, nhit nui v pH mi trng l nhng yu t c bn ti quan trng . C 2 phng php nui VSV: + nui VSV to amylase bng phng php b mt + nui VSV to amylase bng phng php b su Phng php thu nhn t ng vt: Thu nhn amylase t ng vt phn ln l t dch ty tng. 98% ty tng c cu to t t bo ngoi tit hoc l t bo tuyn. Cc t bo ny tit enzyme tiu ha vo trong t trng. VII. Vai tr v ng dng trong cng ngh sn xut ung Trong sn xut bnh m, ngi ta s dng c enzym amylase v amylase cc loi enzym ny thy phn tinh bt to thnh ng. nh nm nen Sacchromysec cerevisiae s d dng chuyn ha chng thnh cn, CO2, lm tng th tch ca bnh v to ra mu sc, hng v tt cho bnh Trong cng ngh sn xut bia ngi ta s dng enzym amylase c trong mm i mch thy phn tinh bt c trong mm i mch. Ngoi ra, ngi ta s dung enzym khc c trong mm i mch chuyn cc cht khng tan sang trng thi tan nh chuyn protein, cellulose, sang aminoacide v glucose. Cc cng ngh ny c thc hin hng ngn nm nay nhiu nc trn th gii. So vi cc enzym amylase t ngun khc, amylase thc i mch ny mm c s dng nhiu nht hin nay. Qu trnh ht i mch ny mm l qu trnh sinh tng hp enzym amylase v nhiu enzym khc. Nh s tng hp ra nhng enzym ny m ht t tin hnh qu trnh t thy phn tinh bt, protein v cc hp cht khc cung cp nng lng cho ht ny mm. qu trnh ny mm l qu trnh trao i cht v l qu trnh sinh l ht chuyn t trng thi nng lng d tr sang trng thi nng lng pht trin

Qu trnh thy phn tinh bt c thc hin theo s sau: Nguyn liu cha tinh bt

Nu chn nhit ng

Amyloglucosidase Hot ng 550C

Lm ngui

Amylase hot ng 550C

ng ha 1. ng dng trong sn xut m chnh: - Nguyn liu s dng ch yu: tinh bt sn, r ng ma. - Cc chng VSV: corymebacterium hydrocacboclastus, bacillus, brevibacterium Enzyme amylase c s dng trong cng on thy phn tinh bt. Ngi ta c th dng a-amylase hay -amylase ca cc ht ny mm hay ca nm mc thy phn tinh bt thnh ng a-amylase Tinh bt Thy phn a-1,4 ca amylose -amylase Tinh bt (glucogen) u im: khng dng n ha cht hay thit b chu acid, chu p lc, khng c hi cho ngi v thit b Nhc im: ng ha khng trit tinh bt m cn dng trung gian nh dextrin lm cho vi khun ln men m maltose + -dextrin. dextrin + maltose + glucose

chnh khng c kh nng s dng. Thi gian ng ha tng i di. Lng ng sau khi ng ha thp, do phi s dng thit b to, cng knh. 2. ng dng trong CNSX bnh m Cc loi ht ng cc nh la m, la nc, bp, khoai m l ngun nguyn liu chnh lm bnh m. Trong sn xut bnh m, ngi ta s dng enzyme nhm gii quyt mt s vn sau: +lm tng nhanh th tch bnh + lm mu sc ca bnh p hn + lm tng mi thm cho bnh Ngi ta s dng c 2 loi enzyme a-amylase v -amylase thy phn tinh bt to thnh ng. Nh nm men saccharomyces cerevisiae s d dng chuyn ha chng thnh cn, CO2, lm tng th tch ca bnh & to mu sc, hng v tt. Ngun amylase thng s dng l malt,tuy nhin trong nhng nm gn y, malt dn c thay th bi nm si. Cc ch phm enzyme t nm si khng ch cha amylase m cn cha protease & 1 s enzyme khc rt c li cho qu trnh ln men. Ch phm enzyme ny ch yu thu nhn t Aspergillus oryzae. Enzyme amylase + sulfat amon hot ng Tc dng: n nh hot tnh enzyme amylase khng thay i trong 1 nm bo qun Tng cht lng bnh (tng th tch, xp, tng kh nng gi hnh bnh, Tng mc n nh cu trc rut bnh & lm gim qu trnh lm kh ca bnh. 1:1 cht m cho amylase

Gim tiu hao nm men lng n 20% Trong cng ngh SX bnh m, lng CO2phi c to ra lin tc t gi u tin n gi cui cng ca qu trnh sn xut. Nu t lng ng c sn trong tinh bt -> CO2 max gi u tin & gi th 2 ca qu trnh ln men sau lng kh gim -> gim cht lng bnh. Khc phc: s dng -amylase -> lng CO2 xy ra theo chiu hng tng & t cc i qu trnh ln men 3. Trong sn xut bnh quy: Tinh bt di tc dng ca enzym Amylase v enzym protelase lm tng hm lng aminoacid t do v tng lng ng kh lm cho bnh quy c mi, v , mu hp dn. 4. Trong sn xut glucose v mt Mt glucoza hay mt maltoza c dng trong sn xut bnh ko v trong cc sn phm n king cho tr em & ngi bnh. Tinh bt di tc dung ca enzym glucoamylase to thnh mt glucoza. Tuy nhin trong ch phm enzyme glucoamylase lun c glucozyltransferaza lm gim tnh cht ca glucoza, lm glucoza kh kt tinh, nh hng xu n cht lng sn phm -> nn chn chng nm mc khng to ra glcozyltranferaza (A.awarnor, Rhizopus,)

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