enzym amylase

8/3/2019 Enzym Amylase

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Enzym Amylase

I. nh ngha v cng th c cu to Amylase l m t henzyme r t phbin trong th gi i sinh v t . Cc

enzyme ny thu c nhm enzyme th y phn, xc tc phn gi i lin k t ni phn

t trong nhm polysaccharide v i s tham gia c a n c:RR + H-OH - > RH + R-OH

C cht tc d ng ca amylase l tinh b t v glycogen. 1. -amylase ( -1,4-glucanohydrolase): (EC 3.2.1.1)

Amylase c kh nng phn ct cc lin k t 1,4-glucoside c a c cht mt cch ng u nhin v lenzyme n i bo. -amylase khng ch c kh nngphn h y htinh b t m cn c kh nng phn hycc h t tinh b t nguyn v n

2. -amylase (glucoamylase) (EC 3.2.1.3)Glucoamylase c kh nng thy phn

lin k t -1,4 l n -1,6-glucoside, ngoi ra

cn c kh nng thy phn lin k t -1,2 v-1,3-glucoside.

Glucoamylase c kh nng thy phnhon ton tinh b t, glucogen, amylopectin,

dextrin thnh glucose m khngcn cs tham gia c a cc lo i enzyme amylase

khc.

3. enzyme isoamylase(glycogen 6- glucanohydolase)(EC 3.2.1.68)


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4. Enzyme pullulanase( -dextrin6- glucosidase)(EC 3.2.1.41)Khc v i dextrinase t i hn trong ho t

ng ca n trn glycogen, v t lcathy phn dextrins gi i hn. Tc ngca n trn Amylopectin hon

tt.Maltose l ng nh nht m n cthchuyn ha m i lin k t.

5. enzyme isoamylase(glycogen 6- glucanohydolase

II. Phn lo i

Enzym chia thnh 2 nhm:

- Endoamylase ( -Amylase)

. -1,4-glucano hydrolase

. -1,6-glucano hydrolase

- Exoamylase

. -amylase

. Amyloglucosidase (glucoamylase)


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III. Nguyn l ho t ng ca enzym

Enzym Amylase l enzym tinh b t, ng vai tr quan trng trong cngnghip thc phm. C chtng qut nh sau:

-Amylase

Dextrin +oligosaccharid

Tinh b t

Dextrin

Pulluulanase

-AmyolaseMaltose

Dextrin

Glucose

Amyloglucosidase

Trong s n xut cn khi th y phn tinh b t, -amylase tc ng vo lin k t -

1,4 glucoside v s n phm to ra mantose v dextrin m m ch ca chng g n

bng C 6. Cc dextrin sau sthy phn ch yu theo:C6 C5 +C 1

C7 C1+ C 6 hay C 5 + C 2

C8 C2+C 6 hay C 3 + C 5

-amylase c a vi khu n thy phn tinh b t tao ra l ng glucose va maltosetheo t l1:5,45. -amylase c a nm s i thy phn tinh b t to ra l ngglucose v maltose theo t l1:3,79. -amylase c a vi khu n v m m s i honton khng c kh nng phn gii -1,6 glucoside c a c cht.trong d ch ha,pH ca dung d ch bt th ng trng h p v i hot ng ti u ca -amylase, pH ny cng l pH hot ng ti u khi tin hnh ng ha. Chnh v th nu


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trong tr ng h p sdng c cc lo i bt c pH trng v i pH ho t ng ti u

ca -amylase th ph i iu ha pH cho thch h pVai tr c a glucoamylase trong s n xut cn: Glucoamylase th y phn lin

k t -1,4 trong cc polysaccharide, chng lin k t ct cc g c glucose khngkh trong m ch polysacchride. Ngoi ra glucoamylase c c kh nng ct link t -1,6 glucoside

Glucoamylase th ng hot ng trong mi tr ng acide (pH ho t ng tiu l khong 3,5-5,5). Ngoi ra cc glucoamylase cn c th hot ng trongmi tr ng trung tnh (pH 6,0-7,5). Cc glucoamylase c a nm men th ng lnhng glucoamylase trung tnh.

Phn l n cc glucoamylase ho t ng pH khng cao. Nhi t ti ukhong 55-60 0C. sn phm cui cng c a hot ng glucoamylase l glucose,glucose c xc nh l ng ln men. Do , vic sdng glucoamylasetrong s n xut cn c tri n vng nh t

Bng. M t s tnh ch t ca glucoamylase

Stt Ngu n enzym Kho ng pH t i u Nhit ti u 1 Aspergillus niger 3,0-6,0

Ti u 4,5-5,055-60

Mt hot tnh 90 0C2 Aspergillus awamori 2,7-7,0

Ti u 3,5-5,055-65

Mt hot tnh 85 0C3 Rhizopus niveus 3,0-6,0

Ti u 3,5-5,0`55-60

Mt hot tnh 70-80 0C

VI. Ngun thu nh n v phng phpthu nh n enzym 1. Ngun thu

ng vt: Enzyme amylase c trong t y tng ca ng vt. Thc vt: c ly tnhiu ngu n nh:

i mch (hodeum sativum):


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+ ging 2 hng: s n xut malt lm bia

+ ging nhi u hng (4,6 hng): lm th c n cho gia sc. La (oryza sativa L.): ch yu vng ng Nam .

Ng (zea mays): c nhi u loi ng: ng , ng bt, ng rng nga.Ht ng c mu tr ng, mu vng hay mu h ng. Ng mu vng do

c shin din ca carotenoid v zeaxanthine trong n i nh. Ngmu h ng l do trong n i nh ca ng c anthocyanin

Vi sinh v t: (VSV) Ngy nay do u thvnhiu mt, vsv tr thnhngun thu enzyme amylase ch o. Nhng chng vsv t o nhiuamylase thng c phn l p tcc ngu n tnhin. Vsv t o

amylase c dng nhi u hn cl nm s i,ginm men v vikhun,cn x khun th t hn.

Cc gi ng nm si th ng dng l gi ng nm s i Aspergillus,rhizopus

Nm men v gi nm men thu c cc gi ng Candida,

Saccharomyces, Endomycopsy, Endomyces cng toamylase.

Nhiu vi khu n c kh nng to l ng ln amylase nh:Bac.polymyxa, Phytomonas destructans, Cassavanumccvi khu n a nhit c kh nng sinh tr ng nhanh v pht tri ntt nhit cao nn khi nui chng t b nhim vsv khc.

Trong nhm x khun rt him gp loi to amylase m nh

m, tuy nhin cng c mt s t nh xkhun a nhit.Micromonospora vugaris 42 c kh nng to mt l ng nh a-amylase ho t ng 65C cng v i protease v cc enzymekhc.


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2. Phng php thu nhn Enzyme amylaseAmylase, m t loi enzym co ngha vmt sinh l, thng mi v

lch s cn c gi l diastase. Enzym ny c c trong th c vt lnng vt. Amylase c tinh s ch t Malt vo nm1835 b i anselmePayen v Jean Persoz.

Amylase thu c nhm Enzym th y phn, xc tc t phn gi i cc lin

k t glucoside n i phn t trong cc polysaccharide v i s tham gia c a

n c. Amylase th y phn tinh b t, glycogen v dextrin thnh glucose,maltose v dextrin h n ch. Amylase c trong n c bt cn c gi lptyalin.

Amylase phn b rng ri trong t nhin v l m t trong nh ng loi

enzym c ng dng rng ri trong cng nghi p. y tv nhi u lnh vckhc, c bit trong cc ngnh cng nghi p thc phm lm l ng tinhbt. Amylase thu nh n tht ny mm, nm mc, nm men, vi khu n.

Trong Amylase c thu nh n tmalt v i s l ng rt nht, chyudng trong s n sut bia

Enzym Glucoamylase. Enzym thu c nhm exo- -1,4D.Glucanglucohydrolate. Theo l thuy t, enzym ny c th y phn tinh b t tothnh glucose c hi u sut n 100% Vi sinh v t: (VSV)

Ngy nay do u thvnhiu mt, vsv tr thnh ngu n thuenzyme amylase ch o. Nhng chng vsv t o nhiu amylasethng c phn l p tcc ngu n tnhin. Vsv t o amylase cdng nhi u hn cl nm s i,ginm men v vi khu n,cn x khunth t hn.


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Cc gi ng nm si th ng dng l gi ng nm s i Aspergillus,rhizopus.

Nm men v gi nm men thu c cc gi ng Candida,

Saccharomyces, Endomycopsy, Endomyces cng to amylase.Nhiu vi khu n c kh nng to l ng ln amylase nh:

Bac.polymyxa, Phytomonas destructans, Cassavanumcc vikhun a nhit c kh nng sinh tr ng nhanh v pht tri n tt nhit cao nn khi nui chng t b nhim vsv khc.

Trong nhm x khun rt him gp loi to amylase m nh m, tuy

nhin cng c mt s t nh xkhun a nhit. Micromonospora

vugaris 42 c kh nng to mt l ng nh a-amylase ho t ng 65C cng v i protease v cc enzyme khc.

Phng php thu nhn malt: Malt i mch:


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Tch m m r

ngm

Thu nh n, x l,

lm s ch, phnS y kh

Ny m m

S y kh

i mch

M m r kh

Malt kh b o

qu n

Bo qu n

Khng kh


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Malt thc:

Phng php tinh sch amylase: 2 phng php. + Tinh s ch enzyme amylase b ng phng php sc k.+ Thu nh n enzyme amylase b ng phng php in di trngel polyacrylamide (polyacrylamide Gel Electrophoresis _

PAGE)

Phng php thu nhn tVSV:S tng h p enzyme amylase khng nh ng ph thuc vo tnh

cht di truy n ca VSV m cn ph thuc vic tuyn chn cc iukin nui c hiu, ngoi ra cc iu kin l ha trong qu trnh nuicng ng vai tr quan trng.

S y malt

Ht thc, laLm s ch, phn

loiS y bo qu n Ngm h t

m mmaltTch m m

malt


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Nhng yu t nh h ng :thnh ph n mi tr ng,tnh ch t c lca mi trng, tit trng, m ban u, thong kh, nhi t nui v pH mi trng l nhng yu t c bn ti quan tr ng .

C 2 phng php nui VSV: + nui VSV t o amylase b ng phng php bmt+ nui VSV t o amylase b ng phng php bsu

Phng php thu nhn t ng vt:Thu nh n amylase t ng vt phn l n l tdch ty tng. 98% t ytng c cu to t tbo ngo i tit hoc l t bo tuy n. Cc t bo ny ti t enzyme tiu ha vo trong t trng.

VII. Vai tr v ng d ng trong cng ngh sn xu t ungTrong s n xut bnh m, ng i ta sdng cenzym amylase v -

amylase cc lo i enzym ny th y phn tinh b t to thnh ng. nh nmnen Sacchromysec cerevisiae s ddng chuy n ha chng thnh c n, CO2,

lm tng thtch c a bnh v t o ra mu s c, hng vtt cho bnhTrong cng ngh sn xut bia ng i ta sdng enzym amylase c

trong m m i mch thy phn tinh b t c trong m m i mch. Ngoi ra,

ng i ta sdung enzym khc c trong m m i mch chuyn cc ch tkhng tan sang tr ng thi tan nh chuyn protein, cell ulose, sang aminoacidev glucose. Cc cng ngh ny c thc hin hng ngn nmnay nhiun c trn th gi i. So v i cc enzym amylase t ngun khc, amylase th c imch ny mm c sdng nhi u nht hin nay.

Qu trnh h t i mch ny mm l qu trnh sinh t ng h p enzym amylasev nhi u enzym khc. Nh s tng h p ra nh ng enzym ny m h t t tinhnh qu trnh t thy phn tinh b t, protein v cc h p cht khc cung c pnng l ng cho h t ny mm. qu trnh n y mm l qu trnh tr ao i cht vl qu trnh sinh l h t chuy n t trng thi nng l ng d trsang tr ng thinng l ng pht tri n


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Qu trnh th y phn tinh b t c th c hin theo s sau:

Nguyn li u cha tinh b t

Nu chn nhit ng

Amyloglucosidase Lm ngu i Amylase ho t ng 55 0C

Hot ng 550C

ng ha

1. ng dng trong s n xut m chnh:- Nguyn li u sdng chyu: tinh b t sn, r ng ma.- Cc ch ng VSV: corymebacterium hydrocacboclastus, bacillus,

brevibacterium Enzyme amylase c sdng trong cng on thy phn tinh b t. Ng i ta c th dng a-amylase hay -amylase c a cc h t ny mmhay ca nm mc thy phn tinh b t thnh ng

a-amylase Tinh b t dextrin + maltose + glucose

Thy phn a-1,4 ca amylose

-amylase Tinh b t maltose + -dextrin.

(glucogen) u im: khng dng n ha ch t hay thi t b chu acid,

chu p l c, khng c hi cho ng i v thi t b Nhc im: ng ha khng tri t tinh b t m cn

dng trung gian nh dextrin lm cho vi khun ln men m


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chnh khng c kh nng sdng. Th i gian ng hatng i di. Lng ng sau khi ng ha th p, do phi sdng thi t b to, cng k nh.

2. ng dng trong CNSX bnh m

Cc lo i ht ng cc nh la m, la n c, bp, khoai m l ngu nnguyn li u chnh lm bnh m.

Trong s n xut bnh m, ng i ta sdng enzyme nh m gii quytmt svn sau:

+lm tng nhanh thtch bnh+ lm mu s c ca bnh p hn

+ lm tng mi thm cho bnh Ng i ta sdng c2 loi enzyme a-amylase v -amylase th y phn

tinh b t to thnh ng. Nh nm men saccharomyces cerevisiae s ddng chuy n ha chng thnh c n, CO 2, lm tng thtch c a bnh& to mu s c, hng vtt.

Ngun amylase th ng sdng l malt,tuy nhin trong nh ng nmgn y, malt dn c thay th b i nm s i. Cc ch phm enzyme t

nm s i khng ch cha amylase m cn ch a protease & 1 s enzymekhc r t c l i cho qu trnh ln men. Ch phm enzyme ny ch yuthu nh n tAspergillus oryzae.

Enzyme amylase + sulfat amon 1:1 cht m cho amylasehot 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 lmkh ca bnh.


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Gim tiu hao nm men lng n 20% Trong cng ngh SX bnh m, lng CO2 phi 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-> CO 2 max gi u tin &gi th 2 ca qu trnh ln men sau lng kh gim-> gim chtlng bnh. Khc phc: s dng -amylase - > lng CO2 xy ra theo chiu hngtng & t cc i qu trnh ln men

3. Trong s n xut bnh quy:

Tinh b t d i tc d ng ca enzym Amylase v enzym protelase lm

tng hm l ng aminoacid t do v tng lng ng kh lm chobnh quy c mi, v , mu h p dn.

4. Trong s n xut glucose v m t

Mt glucoza hay m t maltoza c dng trong s n xut bnh k o vtrong cc s n phm n king cho tr em & ng i bnh.

Tinh b t d i tc dung c a enzym glucoamylase t o thnh m tglucoza. Tuy nhin trong ch phm enzyme glucoamylase lun c

glucozyltransferaza lm gi m tnh ch t ca glucoza, lm glucoza kh

k t tinh, nh h ng xu n cht l ng sn phm -> nn ch n chngnm mc khng t o ra glcozyltranferaza (A.awarnor, Rhizopus,)


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enzym amylase

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