Bao Cao Enzyme Amylase Va Cac Ung Dung

Bo Co Enzyme Amylase & Cc ng Dng
GVHD: TS.Nguyn Cng H
MC LC
I. VAI TR CA ENZYME AMYLASE: 1. Enzyme khc phc khim khuyt t nhin ca nguyn liu...3 2. Enzyme nng cao gi tr thng phm ca nguyn liu..3 3. Enzyme l cng c trong qu trnh chuyn ha cng ngh.5 4. Enzyme tng tnh cht cm quan ca sn phm..6 II. TNG QUAN V ENZYME AMYLASE: 1. Amylase l g? 6 2. Phn loi .8 3. c tnh...9 4. C ch tc dng.12 III. NG DNG CA ENZYME AMYLASE: 1. ng dng ca Enzyme Amylase trong ch bin thc phm gia sc.14 2. ng dng ca Enzyme Amylase trong dc phm...14 3. ng dng ca Enzyme Amylase trong cng nghip dt...15 4. ng dng ca Enzyme Amylase trong vic ty mu giy16 5. ng dng ca Enzyme Amylase trong sn xut bia..18 6. ng dng ca Enzyme Amylase trong sn xut cn.19 7. ng dng ca Enzyme Amylase trong sn xut HFCS20 IV. CNG NGH LN MEN TO ENZYMSE - AMYLASE: 1. Gii thiu v phng php ln men b mt...20 a. u v nhc im ca phng php nui cy b mt..21 i. Mi trng lng..21 ii. Mi trng c...22 b. Qui trnh sn xut nm mc ging23 i. Nguyn liu cho sn xut mc ging.23
Trang 1
http://www.ebook.edu.vn
ii.
Chun b mc ging...23 Nui cy trong ng thch nghing Nui cy ging trong bnh tam gic Nui cy mc trn khay
2. Qui trnh ln men cng nghip to enzym -amylase...25 a. Nguyn liu...25 b. Qui trnh cng ngh...27 c. Thu nhn enzym -amylase t nm mc Aspergillus Oryzae..29 i. ii. Sinh trng v sinh tng hp amylase t nm...29 Cc yu t nh hng n cht lng enzym30 nh hng cc ngun Nit dinh dng nh hng ca Amino acid nh hng ca ngun khong dinh dng iii. Cc yu t nh hng n qu trnh ln men.31
TI LIU THAM KHO .33
I. VAI TR CA ENZYME AMYLASE:
Trang 2
Nguyn liu s dng trong thc phm l cc sn phm nng sn thc t chng l cc vt th sng, do trong qu trnh bo qun hoc ch bin chng thnh cc sn phm thc phm xy ra hng lot cc qu trnh bin i sinh hc m cc qu trnh ny c xc tc t nhin bi enzyme bn th hay do nh cng ngh a vo t mc ch t ra. V vy c th ni enzym ng vai tr ch cht trong cc qu trnh ch bin v bo qun thc phm. Xin gii thiu khi qut 4 vai tr chnh ca enzyme trong C ng ngh thc phm: 1. Enzyme Khc Phc Khim Khuyt T Nhin Ca Nguyn Liu: Cc sn phm nng sn c cht lng v dinh dng, thnh phn ha hc, tnh cht cm quan ph thuc nhiu vo: ging, loi nng sn, iu kin canh tc, iu kin thu hi v vn chuyn, iu kin bo qun. Do vy cht lng sn phm c to ra t nguyn liu dao ng rt ln. Trong thc t nu cht lng nguyn liu qu km ngi ta phi iu khin cc phn ng xc tc bi enzyme to nn cc thnh phn thiu ht trong nguyn liu a vo sn xut. V d: trong sn xut bia, nguyn liu chnh l malt i mch. khc phc malt i mch cht lng km (khng c kh nng chuyn ha ht tinh bt thnh dextrin, ng ln men...) thng dng cc ch phm enzyme thy phn thuc h amilaza nh Termamyl 120L hoc SC , h enzyme proteaza nh Neutrase 0,5L. V d: da , c chua khi a vo sn xut c chn khc nhua, phi qua giai on chn chuyn ha protopectin (1 hp cht pectin c cu to phc tp, ngoi cc chui polygalacturonic cn c cc thnh phn khc nh axit axectic, xenluloza, galactoza, rhamnoza... do chng to nn cu trc cng ca cc loi qu) v dng pectin ha tan nh enzyme pectinaza. Hoc sau khi ch s a enzyme pectinaza vo ph v mng t bo thc vt gip to ra cc vt nt trn qu to thnh rnh thot dch chit ra ngoi lm tng hiu sut chit dch qu. 2. Enzyme Nng Cao Gi Tr Thng Phm Ca Nguyn Liu: Trong thc t c rt nhiu cc nguyn liu nng sn c gi tr thng phm thp. Sau khi c chuyn ha bi tc dng ca enzyme th gi tr thng phm cao hn rt nhiu, chng c th l cc sn phm khng nhng c gi tr dinh dng cao m cn phc v c cho cc mc ch khc ngoi thc phm nh y hc...
Trang 3
V d: Trong cng nghip ch bin tinh bt, mc ch ca nh my ch bin tinh bt l chuyn ha tinh bt t mt hp cht c phn t lng cao, h s hp thu km, gi tr thng phm thp thnh cc sn phm mi c h s hp thu cao hn, a dng hn , gi tr thng phm cao, ng dng nhiu trong cng nghip ch bin cc sn phm khc. Trc y cc nh my ngi ta dng phng php axit: mi trng axit mnh (HCl), nhit cao, p sut cao. Chng c cc nhc im sau: - Phi x l nguyn liu u vo nhm mc ch loi cc thnh phn khc khng pha l tinh bt do axit khng c tnh c hiu. - Sn phm tp, ln nhiu tp cht t thnh phn nguyn liu v t cc sn ng tng hp bi cc thnh phn va c gii phng ra (caramen ha, axit amin+ng to phn ng melanoidin...) lm mu ca sn phm sm, mi kht, nht cao nn qu trnh tinh lc kh khn. - Thit b s dng trong qu trnh ch bin phi c bn cao, chu p sut, chu nhit dn ti gi thnh thit b cao. - Hiu sut thu hi thp dn n gi thnh cao. Vi s pht trin ca ngnh CN enzyme , c bit l CN enzym amilaza th hu ht cc nh my ch bin tinh bt trn th gii chuyn sang s dng phng php enzyme m CN mi ny khc phc oc ton b nhc im trn ca pp axit. Thm vo , trong qu trnh ch bin to ra nhiu sn phm c gi tr cao nh: - Maltodextrin: l cht ph gia ch tc cc sn phm thc phm cho tr em, thc phm truyn trc tip cho bnh nhn. Dng lm tc nhn to cu trc: to c, nht cho nc st; cht kt dnh cho sn xut thc n chnh. Ngoi ra con c kh nng to gel thun nghch c cu trc ging cht bo: thay th cht bo trong cc sn phm b, m gy. - Socbitol: c kh nng gi nc cao to c snh cho mt ong, c ngt nh nn s dng nhiu m khng cm thy kh chu v c nht thp nn d thao tc trong sn xut. Ngoi ra Socbitol cn to phc vi kim loi nng ci thin vic bo qun cc sn phm bo. Socbitol tinh th c th ha tan trong nc rt ln, c kh nng to cm gic mt nn c dng sn xut ko mt, thc n king cho ngi b bnh i ng v khng lm tng lng glucose trong mu sau khi n.
Trang 4
- Manitol: c s dng ch yu lm cht to ngt khng ht nc, c gi tr hp thu thp nn c b sung vo cc loi thc phm chc nng. Bn cnh Manitol c tc ng ph hy rng thp nn c dng lm ko cao su khng ng. 3. Enzyme L Cng C Trong Qu Trnh Chuyn Ha Cng Ngh: Trong cc nh my ch bin thc phm th enzyme c s dng nh mt cng c ca ton b qu trnh chuyn ha hoc l ton b qu trnh chuyn ha trong dy chuyn ch bin. Nu thiu s c mt ca n th qu trnh ch bin khng thnh cng. V d: trong sn xut bia, qu trnh ch bin dch ng c mc tiu l tm iu kin ti u chuyn nhiu nht c th cc cht c trong nguyn liu i vo dch tr thnh cht chit ca dch ng. t c mc ch ny th ngoi qu trnh khuych tn n gin ca cc thnh phn m ch yu l qu trnh chuyn ha cc hp cht cao phn t khng ha tan thnh cc cht thp phn t ha tan di tc dng ca nhiu h enzyme khc nhau, nh: Tinh bt + -Amylase, -Amylase to thnh: dextrin c to snh v gi bt cho bia, maltoza v glucoza: cung cp ng cho qu tnh ln men. Protein + protease to thnh: cc peptit c phn t lng thp lm tng kh nng gi bt do cha nhm chc tch in, to lin kt vi CO2. Cc acid amin dng m bo bn sinh hc do ngn chn s to thnh kt ta to cn ca protein trong qu trnh thanh trng nhit cao do s lm thay i hng s in mi, l ngun cung cp N m bo qu trnh ln men v chng thoi ha ging v lm tng gi tr dinh dng cho bia. Araban (1 hp cht glucan cao phn t lm dch ng rt nht) + arabanase to thnh hp cht thp phn t nn gim nht, ci thin qu trnh lc, tng nng sut dy chuyn. Hp cht cha P + sitase, photphatase lm gii phng P l hp cht quang trng trong chu trnh Kreb v l hp cht vi lng cn cho hot ng sng ca nm men, gip cn bng pH trong mi trng ln men. 4. Enzyme Tng Tnh Cht Cm Quan Ca Sn Phm:
Trang 5
Trong ch bin thc phm sau khi xy ra cc giai on chuyn ha chnh, thng thng cc sn phm thc phm cha hn t cht lng cm quan ti u. V th chng c cc giai on hon thin sn phm , trong giai on ny hoc l to nn nhng iu kin mi cho cc enzyme bn th c li pht huy tc dng c li hoc b sung cc enzyme t ngoi vo lm tng cht lng cm quan ca sn phm nh ci thin mi v v ca sn phm. V d: Qu trnh thy phn lipit nh lipase to thnh hp cht azzetal, xeton c mi c trng ca sn phm. V d: Trong ru vang nh ch yu l pectinase, y l ch tiu cht lng quan trng nht ca ru vang. Vi mc ch lm trong ru cc ch phm enzyme s dng phi c kh nng chu c nng ru r 10-12% v khng cha cc enzyme oxi ha lm nh hng n mu sc v mi v ca sn phm. Hin nay trn th trng c 3 ch phm s dng lm trong dch qu l: Pectinex 3XL, Pectinex AR c th chu c nng ru cao v Viscozyme 120L chu c nng ru thp nhng chu c nng ng cao hnn nn thng c s dng lm trong dch qu c c. Tm li, i vi mi qu trnh sn xut mun s dng cc ch phm enzyme mt cch hiu qu nht ta phi xt y n cc tiu ch la chn nh: hot ng ca enzyme, c hiu ca enzyme v s c mt ca cc cht nh hng n s hot ha v c ch ca Enzyme. II. TNG QUAN V ENZYME AMYLASE: 1. Amylase L G? 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 exoamylase ( enzyme ngoi bo ). - Endoamylase gm c -amylase v nhm enzyme kh nhnh. Nhm enzyme kh nhnh ny c chia thnh 2 loi: Kh trc tip l Pullulanase ( hay RH + ROH C 5 loi enzyme c xp vo 2 nhm: Endoamylase ( enzyme ni bo ) v
Trang 6
dextrin 6glucosidase); kh gin tip l Transglucosylase (hay oligo-1,6-glucosidase) v maylo-1,6-glucosidase. Cc enzyme ny thy phn cc lin kt bn trong ca chui polysaccharide. - Exoamylase gm c -amylase v -amylase. y l nhng enzyme thy phn tinh bt t u khng kh ca chui polysaccharide. C cht tc dng ca amylase l tinh bt v glycogen: Tinh bt: l nhm Carbohydrate thc vt c ch yu trong cc loi c (nh khoai lang, khoai ty, khoai m ), trong cc ht ng cc, cc loi ht v c cng thc tng qut l (C6H12O6)n.Tinh bt t mi ngun khc nhau u c cu to t amylase v amylopectin (Meyer, 1940). Cc loi tinh bt u c 20-30% amylase v 70-80% amylopectin. Trong thc vt, tinh bt c xem l cht d tr nng lng quan trng. Amylase c trng lng phn t 50.000 160.000 Da, c cu to t 200-1000 phn t D-glucose ni vi nhau bi lin kt -1,4-glucoside to thnh mt mch xon di khng phn nhnh. Amylopectin c trng lng phn t 400.000 n hng chc triu Da, c cu to t 600-6000 phn t D-glucose, ni vi nhau bi lin kt 1,4-glucoside v -1,6-glucoside to thnh mch c nhiu nhnh. Tinh bt khng tan trong nc lnh nhng khi hn hp gm nc v tinh bt b un nng (60-850C) th tinh bt s b h ha v c gi l h tinh bt . Di tc dng ca enzyme amylase, tinh bt b thy phn do cc lin kt glucoside b phn ct. S thy phn tinh bt bi enzyme amylase xy ra theo 2 mc : Dch ha v ng ha. Kt qu ca s dch ha l to ra sn phm trung gian dextrin v khi dextrin tip tc b ng ha th sn phm l maltose v glucose. Cabohydrate trong thc phm l ngun cung cp nng lng quan trng cho c th con ngi. Rau v qu cng l ngun cung cp tinh bt v tinh bt ny mt phn c chuyn ha thnh disaccharide v glucose. Carbohydrate c mt trong hu ht cc loi thc phm nhng ngun cung cp ch yu l ng v tinh bt. Glycogen: l mt loi Cabohydrate d tr. ng vt c d tr trong c th ng vt v c c th chuyn ha s dng t t. Amylase c vai tr quan trng trong s chuyn ha glucid t bo ng vt, vi sinh vt. Glucogen c
http://www.ebook.edu.vn Trang 7
cu to t cc glucose lin kt vi nhau bi lin kt -1,4-glycoside cc v tr phn nhnh, glucose ni vi nhau bng lin kt -1,6-glycoside. Glycogen c s mch nhiu hn tinh bt, phn t lng trong khong 2 - 3 triu Da. Glycogen d tan trong nc, nu nh chng ta n qu nhiu Carbohydrate th c th chng ta s chuyn ha chng thnh cht bo d tr. ng vt v ngi, glycogen tp trung ch yu trong gan. 2. Phn Loi: Enzymes Amylase c 5 loi:
Enzym - amylase(EC 3.2.1.1) hay 1-4 glucanohydrolase
Enzyme Pullulanase (EC 3.2.1.41 ) hay - dextrin 6 glucosidase.
Enzyme Isoamylase (EC 3.2.1.68) hay glycogen 6 glucanohydrolase.
Enzyme amylase (EC 3.2.1.2).
Trang 8
Enzyme - amylase (EC 3.2.1.3) hay glucose amylase 3. c Tnh:
Enzyme thy phn tinh bt to thnh dextrin v mt t maltoza. Dextrin c kh nng hat ha cao c trng cho tnh cht ca enzyme ny. Phn t c 1 - 6 nguyn t C, tham gia vo s hnh thnh n nh cu trc bc 3 ca enzyme .Tnh bn nhit ca enzyme - amylase ca sinh vt c nhng c tnh rt c trng v c ch tc ng, chuyn ha tinh bt, kh nng chu nhit: + Th hin hat tnh trong vng axit yu: - Nm mc: pH 4,5 - 4,9 - Vi khun: pH 5,9 - 6,1; pH<3 th enzyme amylase b v hot tr enzyme ca Asp.Niger (pH 2,5 - 2,8). + - Amylase ca nm mc c kh nng dextrin ha cao to ra mt lng ln glucose v maltose. + Nhit ti thch cho hot ng xc tc ca - amylase t cc ngun khc nhau cng khng ng nht. Trong dung dch m pH = 4,7 th - amylase ca Asp. Oryzae rt nhy vi tc ng ca nhit cao, thm ch 400C trong 3 gi th hot lc dextrin ha ca n ch cn 22 - 29% v hot lc ng ha cn 27 - 85%. 500C trong 2 gi th amylase ca nm si ny b v hot hon ton. -Amylase t cc ngun khc nhau c thnh phn amino acid khc nhau, mi loi -amylase c mt t hp amino acid c hiu ring do -amylase l mt protein
Trang 9
giu tyrosine, tryptophan, acid glutamic v aspartic. Cc glutamic acid v aspartic acid chim khong tng lng amino acid cu thnh nn phn t enzyme; amylase c t methionine v c khong 7-10 gc cysteine.Trng lng phn t ca amylase nm mc: 45.000-50.000 Da ( Knir 1956;Fisher,Stein, 1960 ). Amylase d tan trong nc, trong dung dch mui v ru long. Protein ca cc -amylase c tnh acid yu v c tnh cht ca globuline. im ng in nm trong vng pH=4,2-5,7 ( Bernfeld P, 1951 ). -Amylase l mt metaloenzyme. Mi phn t -amylase u c cha 1-30 nguyn t gam Ca/mol, nhng khng t hn 1-6 nguyn t gam/mol Ca tham gia vo s hnh thnh v n nh cu trc bc 3 ca enzyme, duy tr hot ng ca enzyme ( Modolova, 1965 ). Do , Ca cn c vai tr duy tr s tn ti ca enzyme khi b tc ng bi cc tc nhn gy bin tnh v tc ng ca cc enzyme phn gii protein. Nu phn t -amylase b loi b ht Ca th n s hon ton b mt ht kh nng thy phn c cht. -Amylase bn vi nhit hn cc enzyme khc. c tnh ny c l lin quan n hm lng Ca trong phn t v nng Mg2+. Tt c cc amylase u b kim hm bi cc kim loi nng nh Cu2+, Ag+, Hg2+. Mt s kim loi nh :Li+, Na+, Cr3+, Mn2+, Zn2+, Co2+, Sn2+, Cr3+ th khng c nh hng my n -amylase. Thnh phn amino acid ca -amylase nm mc Aspergillus nh sau ( g/100 g protein ): alamine=6,8 ; glycine= 6,6; valine= 6,9; leucine= 8,3; Isoleucine= 5,2; prolin= 4,2; phenylalanine= 4,2; tyrosine=9,5; trytophan= 4,0; xetin= 6,5; trionin= 10,7; cystein v cystine= 1,6; glutamic acid= 6,9; amide= 1,5 ( Akabori et amiloza, 1954 ). Khng ging cc -amylase khc, amylase ca Asp.Oryzae c cha phn phi protein l polysaccharide. Polyose ny bao gm 8 mol maltose, 1 mol glucose, 2 mol hexozamin trn 1 mol enzyme (Akabori et amiloza, 1965). Vai tr ca polyose ny vn cha r, song bit c rng n khng tham gia vo thnh phn ca trung tm hot ng v nm pha trong phn t enzyme. -Amylase ca nm mc hu nh ch tn cng nhng ht tinh bt b thng tn. Sn phm cui cng ca thy phn amylase l glucose v maltose. i vi nm si t l l 1:3,79 (Hanrahan, Caldwell, 1953). -Amylase ca nm si khng tn cng lin kt -1,6 glucoside ca amylopectin, nn khi thy phn n s to thnh cc
Trang 10
dextrin ti hn phn nhnh. y l mt cu trc phn t tinh bt do enzym -amylase phn ct to thnh dextrin ti hn phn nhnh. Sn phm thy phn cui cng ca tinh bt di tc dng ca amylase nm si ch yu l maltose, th n l maltotriose. Khi dng nng -amylase Vi sinh vt tng i ln c th chuyn ha 70-85% tinh bt thnh ng ln men. Cn cc -amylase ca nm mc th mc ng ha n glucose v maltose c th ln ti 84-87%. iu kin hot ng ca -amylase t cc ngun khc nhau thng khng ging nhau, pH ti thch cho hot ng ca -amylase t nm si l 4 - 4,8 (c th hot ng tt trong vng pH 4,5 - 5,8). Theo s liu ca Liphis, pH ti thch cho hot ng dextrin ha v ng ha ca ch phm amylase t Asp.Oryzae trong vng 5,66,2. Cn theo s liu ca Fenixova th pH ti thch cho hot ng dextrin ha ca n l 6,0-7,0. bn i vi tc dng ca acid cng khc khc nhau. -Amylase ca Asp.Oryzae bn vng i vi acid tt hn l -amylase ca malt v vi khun Bac.Subtilis. pH= 3,6 v 00C, -amylase ca malt b v hot hon ton sau 15-30 pht; -amylase vi khun b bt hot n 50%, trong khi hot lc ca -amylase ca nm si hnh nh khng gim bao nhiu (Fenilxova, Rmoshinoi 1989). Trong dung dch -amylase nm si bo qun tt pH 5-5,5; -amylase dextrin ha ca nm si en c th chu c pH t 2,5 - 2,8. 00C v pH= 2,5, n ch b bt hot hon ton sau 1 gi. Nhit ti thch cho hot ng xc tc ca -amylase t cc ngun khc nhau cng khng ng nht, -amylase ca nm si rt nhy cm i vi tc ng nhit. . Nhit ti thch ca n l 500C v b v hot 700C(Kozmina, 1991). Trong dung dch m pH= 4,7 th -amylase ca Asp.Oryzae rt nhy vi tc ng ca nhit cao, thm ch 400C trong 3 gi hot lc dextrin ha ca n ch cn 22-29%, hot lc ng ha cn 27-85%. 500C trong 2 gi, -amylase ca nm si ny b v hot hon ton (Miller v cng s).
Trang 11
4.
C Ch Tc Dng: Giai on dextrin ha:

-amylase
Cc giai on ca qu trnh thy phn tinh bt ca -amylase
Tinh bt Giai on ng ha: Dextrin Amylase Maltose
dextrin phn t lng thp disaccharide&monosaccharide. poliglucose.
tetra v trimaltose oligosacharide maltotriose
maltotetrose.
-Amylase (1,4--glucan-glucanhydrolase) t cc ngun khc nhau c nhiu im rt ging nhau, c kh nng phn cch cc lin kt -1,4-glucoside nm pha bn trong phn t c cht (tinh bt hoc glycogen) mt cch ngu nhin, khng theo mt trt t no c -amylase khng ch thy phn h tinh bt m n thy phn c ht tinh bt nguyn song vi tc t rt chm.
Qu trnh thy phn tinh bt bi -amylase l qu trnh a giai on. + giai on u (giai on dextrin ha):Ch mt s phn t c cht b thy phn to thnh mt lng ln dextrin phn t thp (-dextrin), nht ca h tinh bt gim nhanh (cc amylose v amylopectin u b dch ha nhanh). + Sang giai on 2 ( giai on ng ha ): Cc dextrin phn t thp to thnh b thy phn tip tc to ra cc tetra-trimaltose khng cho mu vi iod. Cc cht ny
b thy phn rt chm bi -amylase cho ti disaccharide v monosaccharide. Di tc dng ca -amylase, amylose b phn gii kh nhanh thnh oligosaccharide gm 6-7 gc glucose(v vy, ngi ta cho rng -amylase lun phn ct amylose thnh tng on 6-7 gc glucopiranose 1). + Sau , cc poliglucose ny b phn cch tip tc to nn cc mch polyglucose colagen c ngn dn v b phn gii chm n maltotetrose v maltotriose v maltose. Qua mt thi gian tc dng di, sn phm thy phn ca amylose cha 13% glucose v 87% maltose. Tc dng ca -amylase ln amylopectin cng xy ra tng t nhng v khng phn ct c lin kt -1,6glycoside ch mch nhnh trong phn t amylopectin nn d c chu tc dng lu th sn phm cui cng, ngoi cc ng ni trn ( 72% maltose v 19% glucose ) cn c dextrin phn t thp v isomaltose 8%. Tm li, di tc dng ca -amylase, tinh bt c th chuyn thnh maltotetrose, maltose, glucose v dextrin phn t thp. Tuy nhin, thng thng amylase ch thy phn tinh bt thnh ch yu l dextrin phn t thp khng cho mu vi Iodine v mt t maltose. Kh nng dextrin ha cao ca -amylase l tnh cht c trng ca n. V vy, ngi ta thng gi loi amylase ny l amylase dextrin ha hay amylase dch ha. III. NG DNG CA ENZYME AMYLASE: Hin nay, vic sn xut ch phm enzyme cc loi a v ang pht trin mnh m trn qui m cng nghip. Thc t c hng nghn ch phm enzyme bn trn th trng th gii, cc ch phm ny a c khai thc v tinh ch c mc tinh khit theo tiu chun cng nghip v ng dng. Ch phm enzyme khng ch c ng dng trong y hc m cn c ng dng trong nhiu lnh vc cng nghip khc nhau, trong nng nghip, trong ha hc " ngha ca vic s dng enzyme trong cc lnh vc thc t khng km so vi ngha ca vic s dng nng lng nguyn t". Theo thi gian, enzym cng nghip ngy cng c ng dng vo nhiu lnh vc khc nhau, trong nhng enzym ng dng nhiu nht l protease, cellulose,
Trang 13
ligase, amylase, v mt s enzym c bit khc. Sau y l mt s ng dng ca enzyme Amylase c s dng trong cc ngnh cng nghip khc nhau. 1. Sc: Trong ch bin thc n gia sc, thnh phn ng cc chim mt khi lng rt ln. Trong khi lng ny, thnh phn tinh bt rt cao. tng hiu sut s dng nng lng t ngun tinh bt, ngi ta thng cho thm enzyme amylase vo. Enzyme amylase s tham gia phn gii tinh bt to thnh ng, gip cho qu trnh chuyn ha tinh bt tt hn. 2. ng Dng Ca Enzyme Amylase Trong Dc Phm. Ngun cung cp enzyme amylase l vi khun Bacillus subtilis vi u im l bn hn trong mi trng acid ca d dy so vi diastase (amylase) ly t ng vt v vi nm. Amylase c s dng phi hp vi coenzyme A, cytocrom C, ATP, carboxylase ch thuc iu tr bnh tim mch, bnh thn kinh. Mt khc amylase phi hp vi enzyme thy phn cha bnh thiu enzyme ng tiu ha. Trong c th, amylase l hormon tuyn tu ngoi tit, c tc dng chng ph n sau chn thng hoc sau m. iu tr triu chng phn ng vim km nhim khun ng h hp trn hoc di. Cc sn phm cha enzyme alpha- amylase dng Alpha-amylase l mt loi enzyme m s kim sot lng calo ca c th. Alpha-amylase l mt men ni sinh c th thy phn tinh bt v cc polysaccharide (hot tnh amylotic) bng cch tn cng vo mi ni a-1,4 glucosidic cho ra cc dextrine, oligosaccharide v monosaccharide. Alpha-amylase dng trong Neopeptine vin nang c ly t ngun vi khun Bacillus subtilis. Amylase ca vi khun cho n nay c u im l bn hn trong mi trng acid ca d dy so vi diastase (amylase) ly t ng vt v vi nm. ng Dng Ca Enzyme Amylase Trong Ch Bin Thc Phm Gia
Trang 14
Sn phm cha men tiu ha dnh cho tr s sinh v tr em, kch thch tiu ha, chng suy dinh dng v cc sn phm cng loi:
MINIGADINE
TRYMO
CALCINOL-RB
CALCINOL 1000
NEOGADINE elixir 150ml
NEOPEPTIN Liquid
3.
ng Dng Ca Enzyme Amylase Trong Cng Nghip Dt
Trong cng nghip dt, ngi ta thng s dng enzym amylase ca vi khun ty tinh bt v lm cho vi mm Trong vi th thng cha khong 5% tinh bt v cc tp cht khc. Do , khi s dng ch phm enzym amylase ca vi khun vi s tt hn. Ngi ta thng s dng lng ch phm amylase khang 0,3-0,6 g/l dung dch v thi gian x l 5-15 pht nhit 900C. Tuy nhin ngoi ch phm enzym amylase c ngun gc t vi khun, hin nay ngi ta quan tm n vic s dng amylase t nm si. Cho n nay c rt nhiu nc s dng enzym trong cng nghip dt tng kh nng cnh tranh cc hng vi, si. Cc nc s dng lng enzym amylase nhiu nht trong lnh vc ny l M, Nht, Php, an Mch. Ngoi ra, enzyme amylase cng c nghin cu ng dng rng ri trong sn xut ng bt, sn xut dextrin, maltodextrin, nha glucose, siro, glucose fructose, sn xut tng v nc chm quy m cng nghip.
4.
ng Dng Ca Enzyme Amylase Trong Vic Ty Mu Giy.
Vic s dng enzyme kh mc giy loi trn th gii c thm d theo nhiu hng v mt s chng vi sinh, theo cc cng b gn y nht cho thy, enzyme -amylase ang c kh nng thc y quy trnh kh mc tt. Nguyn l hot ng ca enzyme l: chng b gy cc mch tinh bt trn mt giy, ko theo s lm long cc phn t mang mu bm trn , to thun li cho qu trnh tuyn ni kh mc. Giai on t nm 2000, Vin Cng nghip giy v xenluyl u t nghin cu mt cch tch cc v s dng enzyme trong cng ngh kh mc giy loi, trong c bit ch trng i vi chng enzyme -amylase. Kt qu nghin cu thnh cng v thit lp quy trnh cng ngh s dng enzyme -amylase kh mc giy loi c gia keo tinh bt. Nguyn liu: giy photocopy, giy vn phng loi, giy in bng mc laser mt mt c gia keo tinh bt. Quy trnh x l: bao gm cc bc ngm v x l giy loi bng enzyme amylase ti nhit thng, x l c ha nhit: nh ti c pha trn ha cht v gia nhit, ti iu kin nhit , pha long v tuyn ni, ra v c c, thnh phm. Ha cht s dng: enzyme -amylase, NaOH cng nghip, Na2SiO3, H2O2, cht hot tnh b mt, cht to bt, du bo. Mu i chng khng s dng enzyme -amylase, cc iu kin cng ngh khc khng i. Hiu qu: Thi gian gim 30%, mc dng cc loi ha cht gim 10 - 35% ty theo tng loi, gim nhiu nht l mc dng NaOH (gim 35%), trng ca bt sau kh mc tng 2,7 - 3,0% ISO, di t tng 1,5%, chu gp (i ln) tng 14% v chu x tng 28%. trng cui cng t ti a 87,0% ISO. Dch v cung cp: Hin nay Vin Cng nghip giy v xenluyl c th cung cp cho cc i tc c nhu cu cc dch v k thut sau: thit k ch to v cung cp thit b ton b dy chuyn x l giy loi bng enzyme -amylase v tuyn ni kh
Trang 16
mc cng sut n 30 tn/ngy, hng dn lp t, chuyn giao cng ngh, o to v hng dn vn hnh. Nm 2008, Vin cng nghip giy v xenluyl tip tc m rng nghin cu cc chng loi enzym khc nhau kh mc giy loi theo Hp ng nghin cu gia Vin vi B Cng thng. Va qua Hp ng nghin cu c nghim thu, trn c s cc kt qu nghin cu c th trin khai ng dng mt s x nghip sn xut bt giy DIP. - Chng loi Enzym s dng: BIO-DE 30 vi thnh phn gm: xylanase, cenllulase v -glucanase, mananase, -amylase vi nguyn liu l giy loi vn phng. - Nguyn liu s dng: Giy loi vn phng - Quy trnh chn la: + Bc 1: x l giy loi bng enzym vi mc dng, pH, nhit v thi gian ph hp. + Bc 2: X l bng ha cht: NaOH cng nghip, Na2SiO3, H2O2, DTPA, cht hot tnh b mt. - Hiu qu thu c: + Tng trng ca bt giy ln 1,53,0 %ISO. + Tng hiu qu loi mc. + Tit kim 1030% ha cht ty theo tng loi. + Gim nhit trong giai on nh ti v 1015oC. + Khng lm thay i ch cng ngh nh s dng phng php kh mc thng thng. + Gim nh hng xu ti mi trng do gim c mc dng ha cht + Cc kt qu ban u cho thy l hon ton kh quan khi tin hnh p dng vo thc t sn xut.
Trang 17
Cng vi vic nghin cu s dng chng loi enzym BIO-DE 30, Vin cng giy v xenluyl ang tip tc nghin cu ng dng mt s loi chng loi enzym th h mi, s dng cho qu trnh kh mc giy bo v tp ch. - Dch v cung cp: Thit k ch to h thng thit b tuyn ni, hng dn lp t, chy th v giao cng ngh kh mc giy loi vn phng s dng enzym BIO-DE 30 kt hp vi s dng ha cht. 5. ng Dng Ca Enzyme Amylase Trong Sn Xut Bia: Trong cng ngh sn xut bia truyn thng, cc nc phng Ty ch yu s dng enzym amylase ca malt thy phn tinh bt trong malt, sau n giai on ru ha bi nm men Saccharomyces sp. C s khoa hc ca vic s dng amylase ca malt ch, khi i mch chuyn t trng thi ht sang trng thi ny mm (malt), enzym amylase s c tng hp v khi enzym ny s thy phn tinh bt c trong ht to ra nng lng v vt cht cho s to thnh mm. Nh vy vic ng ha tinh bt trong ht nh enzym ca chnh n. Khi ht ch tng hp ra lng enzym amylase va phn hy lng tinh bt c trong ht. Nh th cn rt nhiu mm i mch sn xut bia qui m ln, dn n chi ph cao cho sn xut v sn phm. khc phc iu ny, trong qu trnh ln men to bia th nh sn xut khng s dng hon tan 100% nguyn liu l malt i mch m c s pha trn theo mt cng thc no thay th malt v cn b sung ngun tinh bt cho qu trnh ln men. L do l mt phn to hng v cho bia, mu sc, cn ph hp cho ngi tiu dng v mt phn l lm gim gi thnh cho sn phm bia nhng vn gi c c trng cho bia. Chnh v iu ny, cc nh sn xut bia quan tm n vic s dng ch phm enzym amylase cung cp cho qu trnh thy phn tinh bt .Enzym ny c ngha rt ln trong vic lm bia, gip sn xut bia qui m cng nghip.
Trang 18
Ngoi ra, trong sn xut bia, ngi ta cn s dng ch phm enzym cellulose c tc dng ph v thnh t bo, to iu kin cc thnh phn c trong t bo ht that ra pha ngoi nh cht lng bia c nng cao hn. Mt loi enzym khc cng c s dng kh rng ri l gluco amylase, enzym ny c s dng loi tr O2 c trong bia, gip qu trnh bo qun bia ko di hn rt nhiu. 6. ng Dng Ca Enzyme Amylase Trong Sn Xut Cn: sn xut cn t ngun nguyn liu tinh bt, mi nc s dng cc loi nguyn liu khc nhau.V d, M ngi ta s dng nguyn liu t bt ng sn xut cn, cn Brazin li s dng khoai m, cc nc khc s dng go hoc tm t go. Qu trnh sn xut cn tri qua hai giai an: giai an ng ha v giai an ru ha. Giai an ng ha, ngi ta bt buc phi s dng enzym amylase (khng th s dng phng php thy phn tinh bt bng acid). Ngi Nht bit s dng enzym ca nm mc trong qu trnh ng ha sn xut ru Sake t cch y hn 1700 nm. Ngi Trung Quc th s dng nhiu loi nm mc ng ha ru trong sn xut ru cch y 4000 nm. Cn ngi Vit Nam bit sn xut ru t go cch y hng ngn nm. Ring M, mi n th k XIX khi Takamine ngi Nht a nm mc Aspergillus sang mi bit s dng enzym ny thay amylase ca malt sn xut cn. Chnh v th mi c phng php Micomalt ( mm mc) trong sn xut cn v ru. Nh s du nhp k thut ny t Nht m ngi M tit kim c mt khi lng malt khng l trong sn xut ru. Giai an ru ha, nh nm men Saccharomyces cerevisiae, cng c th xem y l mt qu trnh p dng enzym. Qa trnh ru ha l qu trnh ht sc phc tp, tri qua rt nhiu giai on chuyn ha t ng thnh cn nh s tham gia ca nhiu enzym khc nhau. im khc vi enzym amylase l ch cc enzym tham gia qu trnh ru ha nm trong t bo nm men. Vic iu khin cc qu trnh chuyn ha
Trang 19
bi enzym trong t bo thc cht l qu trnh trao i cht ca nm men trong mi trng cha ng. 7. ng Dng Ca Enzyme Amylase Trong Sn Xut HFCS: HFCS (High Fructose Corn Syrup) l mt nhm bt k ca siro bp m tri qua qu trnh enzyme lm tng hm lng fructose v sau c pha trn vi siro bp sch (100% glucose) to thnh dng sn phm cn thit. Nguyn nhn chnh l: + Nguyn liu r M, n ph thuc vo gi c ca bp v ng. + Sn phn d dng pha trn v di chuyn bi v n dng lng. Cc dng ph bin ca HFCS: + HFCS 90 + HFCS 55 + HFCS 42 Qu trnh enzyme lm thay i 100% glucose siro bp thnh HFCS 90 theo cc bc sau: + Tinh bt bp c x l bng -amylase sn xut chui ng ngn hn gi l oligosaccharides. + Glucoseamylase ph v nhng chui ng sn xut ng n glucose. + Xylose isomerase bin i glucose to ra hn hp c khong 42% fructose v 50- 52% glucose vi mt vi loi ng c ha trn. + Hn hp 42 - 43% fructose glucose phi chu qua mt bc sc k lng ni m fructose s c lm giu n xp x 90%. IV. CNG NGH LN MEN TO ENZYMSE - AMYLASE: 1. Gii Thiu V Phng Php Ln Men B Mt: Cui th k 19 u th k 20, vic nui cy Vi sinh vt thng c thc hin theo phng php b mt. Phng php ny c pht trin rt rng ri, khng ch
Trang 20
thu nhn ch phm enzyme m trc tin l phng php thu nhn khng sinh v mt s qu trnh ln men truyn thng. a. u v nhc im ca phng php nui cy b mt: Phng php nui cy b mt l phng php to iu kin cho Vi sinh vt pht trin trn b mt mi trng. Nhng u im ca phng php nui cy b mt l: - Nui cy b mt rt d thc hin. Quy trnh cng ngh thng khng phc tp. - Lng enzyme c to thnh t nui cy b mt thng cao hn rt nhiu so vi nui cy chm. y l c im u vit rt quan trng trong gii thch ti sao phng php nui cy b mt hin nay pht trin mnh tr li. - Ch phm enzyme th (bao gm thnh phn mi trng sinh khi Vi sinh vt, enzyme v nc). Sau khi thu nhn rt d sy kh v d bo qun. - Nui cy b mt khng cn s dng nhiu thit b phc tp, do vic vn hnh cng ngh cng nh vic u t va n gin va khng tn km. - Trong trng hp b nhim cc Vi sinh vt l, ta rt d dng x l. Mi trng c l mi trng tnh, khng c s xo trn nn khu vc no b nhim ta ch cn loi b khu vc khi ton b khi nui cy. Nhng khu vc khc s hon ton c an ton. Phng php nui cy b mt cng c nhng u im cn quan tm khc phc v hon thin dn phng php ny. Nhc im ln nht v d nhn thy nht l: Phng php ny tn kh ln din tch cho nui cy. Trong phng php ny Vi sinh vt pht trin trn b mt mi trng (mi trng lng hoc mi trng bn rn) nn rt cn nhiu din tch. i. Mi trng lng: Trong mi trng lng, Vi sinh vt s pht trin trn b mt mi trng, to thnh khun lc ngn cch pha lng (mi trng) v pha kh (khng kh). y, Vi sinh vt s s dng cht dinh dng t dung dch mi trng, O2 t khng kh, tin hnh qu trnh tng hp enzyme. Enzyme ngoi bo s c tch ra t sinh khi v ha tan vo dung dch mi trng. Enzyme ni bo s nm trong sinh khi Vi sinh vt.
Trang 21
Nui cy b mt trn mi trng lng trong cc khay: Nui cy Vi sinh vt thu nhn enzyme trn mi trng lng theo phng php cy b mt thng c tin hnh trong cc khay c chiu cao khong 12-15 cm, chiu rng v chiu di c thit k ty theo kch thc phng nui sao cho thun tin trong thao tc. y ngi ta quan tm nhiu n chiu cao mi trng lng. Nu chiu cao mi trng lng qu ln, Vi sinh vt s khng c kh nng ng ha ht cc cht dinh dng pha y khay nui cy. Nu chiu cao mi trng nh, s thiu thnh phn cht dinh dng, hiu sut thu nhn enzyme s khng cao. Trong nghiu nh my, ngi ta thng to mi trng trong khy nui cy c chiu cao mi trng t 5-7 cm l hp l. ii. Mi trng c: Phn ln cc nh my sn xut enzyme, khi nui cy Vi sinh vt thu nhn enzyme, ngi ta thng s dng mi trng c . tng kh nng xm nhp ca khng kh vo trong lng mi trng, ngi ta thng s dng cm, tru, ht ng cc lm mi trng. Trong trng hp ny, Vi sinh vt pht trin trn b mt mi trng, nhn cht dinh dng t ht mi trng v sinh tng hp ra enzyme ni bo v ngoi bo. Cc enzyme ngoi bo s thm thu vo trong cc ht mi trng, cn cc enzyme ni bo nm trong sinh khi Vi sinh vt. Vi sinh vt khng ch ph trin trn b mt mi trng, ni ngn cch pha rn (mi trng) v pha kh (khng kh) m cn pht trin trn b mt ca cc ht mi trng nm hn trong lng mi trng. Mi trng nui cy va c xp cao v va phi c m thch hp. Nu m qu cao s lm bt mi trng li , khng kh khng th xm nhp vo trong lng mi trng, nu c m thp qu s khng thun li cho Vi sinh vt pht trin. Thng thng ngi ta thng to m khong 55-65% W l hp l. Nu s dng cm lm nguyn liu chnh nui cu Vi sinh vt thu nhn enzyme, ngi ta phi cho thm 20-25% tru lm xp mi trng, to iu kin thun li khng kh d xm nhp vo lng mi trng. Phng php nui cy b mt bn rn(mi trng c ) ny rt thch hp cho len men nm mc Asp.oryzae.
Trang 22
b. Qui trnh sn xut nm mc ging: i. Nguyn liu cho sn xut mc ging: Go np:Nc 14% ,Glucid 79,4%, Lipid 1,5% v protein 8,2% trong Protein ca go np ch yu l glutein(oryzaine)v glubuline ngoi ra cn mt t ln Cozine v Prolamin. Glucid ca go np ch yu l tinh bt, ng, cellulose v hemicellulose.Trong tinh bt ch yu l amylopectin.Ngoi ra cn cha mt s vitamin nh B1,B2,B6,PP v E. Go t:Nc 13,84%; Glucid 77,55%; Protein 7,35%; Lipid 0,52%; cellulose 0,18% v mui khong :0,54%. Bt M:Nc 11,6%; Glucid 72%; Lipid 4,8%,Protein 9%; cellulose 1,5% v mui khong 1,2%. Bp mnh:Nc 11,4%; Glucid 78,9%; Lipid0,8%; Protein8,5%,cellulose 0,4% v mui khong 0,4%. ii. Chun b mc ging: Nui cy ging bao gm: - Trong ng thch nghing hay gi ging trong ng nghim. - Trong bnh tam gic (nhn ging nh). - Trn sng, khay (nhn ging ln). Nui cy trong ng thch nghing: Sau khi phn lp thnh cng trn mi trng chn lc t a petri cy chuyn nhng mc si sang (hay ly nm mc t ng ging) ng thch khc.Yu cu ng ging phi tuyt i m bo thun khit, khng c ln ln bt k mt loi Vi sinh vt no khc.Mi trng thch nghing phi m bo y dinh dng. Nui cy ging trong bnh tam gic: Cch lm mi trng trong bnh tam gic: Mi trng go:go t loi tt, nu cm nh nu bnh thng, ht cm chn u khng qa nho v khng qa kh. m khong 45% ngui bp ri
Trang 23
thnh tng ht cho vo bnh tam gic thnh lp dy khong 1cm. y nt bnh tam gic bng giy chng m. Hp thanh trng P = 1atm trong vng 30-45 pht. Mi trng ng mnh: Ng mnh c kch thc khong 0,2-0,5 mm cho nc vo theo t l 90% trng lng so vi ng, trn u trong khay 1-2 gi cho ngm nc u. Bp ti cho vo bnh tam gic khc thnh lp dy 1cm. y nt bnh bng giy chng m. Hp thanh trng ng thi lm chn P = 1atm, 1200C, trong thi gian 60 pht, ly ra ngui lc cho khi vn cc. Mi trng cm: Chn cm tt, mi nhng loi th khng cn mn ht, sau lm tng t nh i vi ng mnh. Thng s dng cc bnh tam gic dung tch 0,3-0,5 lt hay 1 lt c c rng. Sau khi chun b mi ttrng trong bnh thu tinh ta tin hnh nui cy nm mc. Trc tin cn phi chun b ly 5ml nc v trng cho vo cc ng nghim. Sau , nc v trng cho bo t ho vo trong nc ng thi cy chuyn chng sang bnh tam gic. Trung bnh c mt ng nghim c th cy chuyn sang 2-3 bnh tam gic c dung tch 1 lt, lc cho ging phn b u trong mi trng v tin hnh nui cy chng trong iu kin thch ng. Thng nui khong 5-6 ngy l c. Yu cu c bn trong giai on ny l lm sao to c nhiu bo t mnh kho. Trng hp no thy bnh b nhim th phi loi b ngay. Nui cy mc trn khay: Chun b mi trng lm mc trn khay: Nguyn liu thng dng l ng mnh c kch thc 0,2-0,5mm. Trn vi nc theo t l 80-90% trng lng so vi ng nu hp di p lc cao. Trn xong khong 3-4 gi cho ng ngm nc u ri hp chn. Thi gian hp khong 3-4 gi, di hn so vi thi gian hp cm hay go. Nguyn liu sau khi hp phi chn u, khng c qu bt hoc qu kh. m cn li 45-50% l va. Cch gieo cy v nui mc trn khay: Nguyn liu d ra, lm ngui nhanh. Nu t c th bp bng tay cho ti ra, nu nhiu cho qua my nh ti v dng qut thi. Sau khi lm ngui n 26-280C th trn nc ging t bnh tam gic vo vi t l 0,5-1% hoc vi t l cao hn.
Trang 24
Sau khi trn ging, mi trng vo khay thnh lung cao (0,3m). t nhit 30-320C, m 85-100%. Thi gian 6-8 gi, nhit khi mi trng ln ti 34-360C v bo t ny mm gn ht nhng cha thnh si di. Lc ny cn ri mi trng thnh lp mng 2-3cm cn gi cho nhit mi trng khng vt qu 360C. ng nhit ta ra nhiu nht khong 10-14 gi sau khi trn ging. Sau khi nui 34-36 gi, nhit khi mi trng bt u gim, cn phi iu chnh nhit ln 34-350C, duy tr s hnh thnh bo t ca nm mc. Thi gian nui mc ging trn khay thng vo khong 60 gi. Nu thy hnh thnh bo t chm th c th ko di n 70-72 gi. Mc ging khi ly ra thng c m 32-35 % c th dng ngay lm ging cho cng on sn xut. Nu khng dng ngay th phi em sy kh n m 8%, gi dng dn hoc l cung cp ging cho cc ni sn xut. Nhit phng sy khng c qu 400C. Cc bao mc ging cn c bo qun ni thong, mt ( c th bo qun lnh 4-50C ) trnh nh nng. Thi gian bo qun ty iu kin c th, t 1-2 thng hoc lu hn. 2. Qui Trnh Ln Men Cng Nghip To Enzym lpha-Amylase: a. Nguyn liu: Ngun tinh bt: nc ta ngun nguyn liu cha tinh bt th v cng a dng v phong ph. V d tinh bt t go (go t, go np, tm), ng, sn, Nhng ph hp cho sn xut enzym amylase qui m cng nghip th cn phi tnh n chi ph cho gi thnh sn phm. Do vy, ngun nguyn liu cn c quan tm v ch trng v mc r tin v d kim. y l mt li th rt quan trng cho nh sn xut v cho c ngi tiu dng c gi thnh thp v c sn xut i tr. Sau y, gii thiu mt s ngun nguyn liu d tm: - Thnh phn dinh dng ca ht go: Thnh phn dinh dng /100g go: Protein 6g, Tinh bt 82g, Lipid 0.8g,Cellulse 0.6g, Nc 10.2g, Nng lng 361 kcal (Vin Nghin cu dinh dng thuc i hc Mahidol, Thi Lan, 1999). Protein ca go chim khong 8-9%, ch yu l glutelin v glubuline. Ngoi ra cn cln t : cozine v prolamin. Lng protein thoi ho v bin tnh dn trong qu trnh bo qun. Glucid th ch yu l tinh bt, ng, cellulose hemicellulose.Trong tinh bt
Trang 25
ch yu l amylopectin, c cha mt t cht khong nh: P, K, Mg, Ngoi ra cn c mt s Vitamin: B1, B2, B6, PP, H, C, E v carotenoid. - Thnh phn dinh dng ca ng: Thnh phn dinh dng /100g bp ngt: Protein 9.1-12.9, Tinh bt 69.3-72.8, Lipid 2.2- 3.9, Cellulose 1.8-2.9, Nc 9.5-11.2, Khong 1.5-2.5 (Cortez v Wild-Altamirano,1972). - Thnh phn dinh dng ca sn (khoai m): Sn l loi c cha nhiu tinh bt, rt thch hp l ngun c cht cm ng cho qu trnh tng hp enzym amylase nm mc Asp.oryzae. C sn gm ba phn chnh: v, tht c v li. Ngoi ra cn c cung v r c. + V gm hai phn: V g bn ngoi, cu to ch yu l cellulose, thng chim khong 1,5-2% khi lng c, v ci cng cu to t cellulose nhng trong v ci cn c m sn l cc polyphenol, chim ti 85-90% polyphenol ca c sn. + Tht cha nhiu tinh bt, t protein v mt lng du, lng polyphenol y c khong 10-15%, nhng cc polyphenol gy tr ngi khi ch bin, t bit l sn chy m s lm cho bt sn bin mu, thay i mi v kh n trc tip khi luc, kh thot nc khi sy hay phi kh sn lt hay sn bt. + Thnh phn ho hc ti ca sn: Tinh bt 20-34%; protein 0,81,2%; cht bo 0,3-0,4%; cellulose 1-3,1%; cht tro 0.54%; polyphenol 0,1-0,3%; nc 60-74,2% . Nguyn liu s dng trong nui cy b mt thng l nhng nguyn liu c ngun gc t nhin nh cm m, cm go, go, ng mnh, u nnh v cc loi ht ng cc khc.Trong cc loi nguyn liu trn, cm go, cm m c s dng nhiu hn c. Hai loi ny c y cc cht dinh dng cn thit cho Vi sinh vt pht trin. Mt khc khi to mi trng, chng thng c tnh cht vt l rt thch hp va m bo khi kt dnh cn thit, va m bo lng khng kh lu chuyn trong khi nguyn liu. Trong nhiu trng hp, to kh nng thong kh tt hn, ngi ta thng cho thm tru vi t l thch hp cho tng loi enzym c to ra t Vi sinh vt. Thc cht cho tru vo l lm tng xp ca mi trng, to nn nhng khong trng khng kh c th lu thng trong lng mi trng. Chnh v th ta thy rng
Trang 26
nm mc Asp. oryzae khng ch pht trin trn b mt mi trng m cn pht trin rt mnh trn b mt ht mi trng. Hay ni cch khc, chng nm mc Asp.oryzae c kh nng pht trin gia hai pha rn v pha kh ca mi trng.Trong trng hp ny, n c kh nng pht trin hn trong lng mi trng nhng n vn hon ton mang ngha ca qu trnh ln men b mt. b. Qui trnh cng ngh: Nguyn liu: Trong phng php ln men b mt, cu trc chnh l cm m, cm go. Hai loi ny l nguyn liu hon ho v c th l mt cu t duy nht ca mi trng nui nm khng cn b xung thm cht khc na. Tuy nhin do l ph liu ca cng ngh xay xt, nhng lm cng tng i t tin. Hn na trong qu trnh nui nm, cc cht dinh dng khng c s dng ht, v th c th thay th hoc pha trn thm mt s cu t r tin hn. Mt khc cu t b sung vo (tru, mt ca,) lm tng xp gip cho h si nm pht trin tt hn tn dng ti a ngun c cht sinh ra hot lc enzyme cao nht. c bit ch , cm khng c cha tinh bt di 20-30%. Nn dng cm tt, cm mi khng c v chua hay ng, khng hi mi mc. m ca cm khng qu 15%, tp cht c khng qu 0,05%. Hp thanh trng: Cm v cc cht ph gia cha nhiu bo t Vi sinh vt khc nn cn phi thanh trng m bo chng nui pht trin bnh thng v canh trng sn xut khng cha Vi sinh vt ngoi lai, cn thanh trng di p sut hi 11,5 atm trong thi gian 45-60 pht. thun li cho vic thanh trng c hiu qu, cng nh to iu kin cho nm si pht trin, trc khi thanh trng, ngi ta thng dng HCl hay H2SO4 iu chnh pH mi trng. Sau khi thanh trng mi trng c lm ngui ngi ta bt u nui cy thu nhn enzyme. Trn ging vi sinh vt: Sau khi lm ngui, tin hnh cy ging hoc rc bo t vo mi trng thanh trng, thnh ng vi gi, ging mc c th nui cy ring phng th nghim hay ti cc phn xng nhn ging hoc l ging thng phm c bn ni cung cp ging. Cc loi ging ny thng cha nhiu bo t. Khi cy vo mi trng dinh dng, chng s pht trin thnh t bo nm mcv to ra cc loi enzyme m ta mong mun.T l ging a vo nui cy thng vo khong 0,5-20% so vi khi lng ca mi trng.
Trang 27
K thut nui cy: Sau khi trn ging, mi trng c tri u ra cc khay vi chiu di 2-3cm, ri c a vo phng nui cy, t trn nhng gi . Cc gi ny c thit k sao cho lng khng kh c lu thng thng xuyn. Phng nui cy phi c h thng iu chnh nhit v m khng kh.Nhit thch hp cho nm si pht trin l 28-320C. Nhit thp qu hoc cao qu u nh hng khng tt cho nm si pht trin. Trong qu trnh nui cy, ta hon ton khng cn iu chnh pH. Mi trng bn rn l mi trng tnh nn s thay i pH mt vng no t khi nh hng n ton b khi mi trng. Thi gian nui nm si thu nhn enzyme vo khong 36-60 gi. iu ny cn ph thuc vo chng nm mc Asp.oryzae v iu kin mi trng cng nh ph thuc vo iu kin nui cy. Qu trnh pht trin ca nm mc trong mi trng bn rn khi nui bng phng php b mt ny tri qua cc giai on sau: Giai on 1: Giai on ny ko di 10-14 gi k t thi gian bt u + Nhit tng rt chm. + Si nm bt u hnh thnh v c mu trng hoc mu sa. + Thnh phn dinh dng bt u c s thay i. + Khi mi trng cn ri rc. + Enzyme mi bt u c hnh thnh. Trong giai on ny phi c bit quan tm n ch nhit . Tuyt i khng c a nhit cao qu 300C v thi k u ny ging rt mn cm vi nhit . - Giai on 2: Giai on ny ko di 14-18 gi. Trong giai on ny c nhng thay i c bn sau: Ton b bo t pht trin thnh si nm v si nm bt u pht trin rt mnh cc si nm ny to ra nhng mng si chng cht khp trong cc ht mi trng trong lng mi trng. Trong giai on ny ta c th hon ton nhn r cc si nm c mu trng xm bng mt thng. + Mi trng c kt li kh cht. + m mi trng gim dn. nui cy. giai on ny c nhng thay i sau:
Trang 28
+ Nhit mi trng s tng nhanh c th ln ti 40-450C. + Cc cht dinh dng bt u gim nhanh do s ng ho mnh ca nm si. + Enzyme amylase c tng hp mnh. + Lng O2 trong khng kh gim v CO2 s tng dn, do trong giai on ny cn phi c thng kh mnh v nhit c gng duy tr trong khong 29300C l tt nht. - Giai on 3: Giai on ny ko di 10-20 gi. giai on ny c mt s thay i c bn nh sau: + Qu trnh trao i cht yu dn, do mc gim cht dinh dng s chm li. + Nhit ca khi mi trng gim, do lm gim lng khng kh mi trng xung 20-25 th tch khng kh /th tch phng nui cy/ 1gi. Nhit d nui duy tr 300C,trong giai on ny, bo t c hnh thnh nhiu do lung enzym amylase to ra s gim xung . Chnh v th vic xc nh thi im cn thit thu nhn enzym rt cn thit. c. Thu nhn enzym -amylase t nm mc Aspergillus Oryzae: i. Sinh trng v sinh tng hp amylase t nm: Khi nui Vi sinh vt to amylase c 2 qu trnh lin quan mt thit vi nhau. Qu trnh tng hp sinh khi Vi sinh vt v qu trnh tch t enzyme trong t bo hoc ngoi mi trng. mt s Vi sinh vt, qu trnh sinh tng hp amylase tin hnh song song vi qu trnh sinh trng, ngha l s tch t enzyme ph thuc tuyn tnh vo s tng khi. Trong trng hp ny, sinh tng hp enzyme amylase kt thc pha logarit cng ng thi vi s ngng sinh trng v s bt u pha pht trin n nh tip theo sau. Tuy nhin s to thnh amylase cc i thng xy ra sau khi qun th t bo Vi sinh vt t im sinh trng. Trong trng hp ny, sinh trng ca Vi sinh vt hu nh khng km theo s tch ly enzyme amylase trong canh trng, ch sau khi kt thc pha sinh trng mi xy ra s tng hp enzyme cc ln.
Trang 29
ii.
Cc yu t nh hng n cht lng enzym: nh hng cc ngun Nit dinh dng:
Ngun dinh dng Nit ht sc quan trng. Khi chun b mi trng dinh dng nui nm mc Asp.oryzae to enzym -amylase, ngi ta dng mui v c Natri Nitrat l ngun Nit dinh dng nui nhiu loi nm si to amylase. Hm lng thng s dng l 0,91%. Nari Nitrat v Amoni Nitrat c hiu qa hn so vi cc loi mui khc ( Kali Nitrat, Magie Nitrat, Amoni Sunfat.). Cho ngun Nit nht nh vo mi trng c th kch thch tng hp amylase ny v c ch tng hp amylase khc .V d:Kt qa th nghim Fenikvova v cng tc vin v vic s dng cc ngun Nit t nhng mui khc nhau. Ngoi ra cng cn phi xc nh t l gia ngun carbon v Nit trong mi trng ph hp cho mc Asp.oryzae pht trin. Tnh cn bng ca mi trng dinh dng v carbon v Nit c ngha ln i vi s sinh tng hp sinh khi ca nm Asp.oryzae v s to thnh amylase. nh hng ca Amino acid: Amino acid l nhng cu t hp thnh phn t enzym, trong c mt s cc amino acid khng ng nht v gi tr dinh dng nn s dng hn hp amino acid s c gi tr ln hn cho nhng cht lng mi. Amino acid c nh hng tt ti sinh l ca nm mc cng nh vic sinh tng hp enzym amylase do nhng nguyn nhn sau: - Amino acid c th ng thi va l ngun carbon , ngun Nit va l ngun nng lng. - Mt s amino acid ring l (Glutamic acid ,Aspatic acid.) ng vai tr v cng quan trng trong trao i amino acid c th l sinh tng hp nhiu amino acid khc trong qa trnh chuyn amine ha. Nh mt s amino acid trc ht l glutamic acid, aspatic acid m thc hin ng ha d dng CO2 (Araiet al,1968). - Nhiu amino acid tham gia vo thnh phn ca cc vitamin tan trong nc nh: Pantotenic acid, biotin, folic acid, cc vitamin ny c ngha sinh hc rt ln. Ngoi ra cn c nhng nhn xt cho rng s tng hp ARN trong t bo mt s Vi sinh vt ph thuc vo s c mt ca amino acid trong mi trng nui ( Kurland, Marloe, 1962 ).
Trang 30
- Amino acid c tnh c hiu ca s cm ng v kim ch sinh tng hp enzyme. Mt s amino acid kch thch v lm tng cng sinh tng hp cc enzyme amylase, mt s li c tc dng c ch ngc li v mt s khc th khng c nh hng g c. nh hng ca ngun khong dinh dng: Cc yu t a lng v vi lng c nh hng ln n sinh trng v tng hp cc enzym, -amylsae nm mc Asp.oryzae. - Ion Mg2+ c nh hng ti bn nhit ca enzym. Thiu MgSO4 s c nh hng xu n s tng hp mi amylase bi nm si. Khi s tng hp amylase b c ch hon ton (Fenikxova, Muxaeva, 1967). Nng ti u ca mui ny cho tng hp -amylase l 0,05%. - Phospho cn tng hp cc hp phn quan trng ca sinh cht (nucleic phospholipide acid) v nhiu coenzym, ng thi phosphoril ha glucide trong qu trnh oxy ha sinh hc. Phospho nh hng trc tip ti sinh sn ca nm si, do vy m tng cng tng hp enzym. Cc mui phosphate thng c dng vi nng cao ti 0.15M. Hot ca -amylase tng cao khi c 0,1% KH2PO4. - Canxi (Ca) cn cho tng hp v n nh -amylase hot ng v n l cu t khng th thiu c ca enzym ny. Canxi cn c tc dng bo v amylase khi tc ng ca protease (Hsiu et amiloza, 1964). Mun tch ly nhiu -amylase cn c lng Ca trong mi trng l 0,01-0,05%. - Lu hunh (S) kch thch s to amylase ( S c trong thnh phn ca cc amino acid quan trng : methionine, cystein, cystine). Lu hunh vi hm lng 0.04g/ml mi trng l thch hp nht cho Asp.oryzae to amylase. Ngoi nhng nguyn t a lng ny cn ch thm vi nhng nguyn t vi lng. Chnng hn nh: Coban, km,v cc nguyn t gy c ch s tng hp enzym amylase nh: ng, thy ngn, iii. Cc yu t nh hng n qu trnh ln men: m(w) mi trng: Tt nht cho s hnh thnh enzym ca nm mc Asp.oryzae l 55-60%. m mi trng thch hp cho s hnh thnh bo t l khong 45% nn cn gi cho m mi trng khng b gim trong qu trnh pht
Trang 31
trin. nh hng ca vic gi m trong qu trnh sinh trng ti s to -amylase ca Asp. oryzae nui bng phng php b mt: iu ny tht s cn thit phi gi m cho mi trng mc ti thch. Cn thng kh lin tc trong sut thi k sinh trng ca nm mc v cn ph thuc vo kch thc, chiu dy ca lp mi trng nui. - m tng i ca khng kh: m cho php t 80% tr ln n mc bo ho tc l 100% W u thch hp cho nm mc pht trin. Trong phng nui cn gi cho m khng kh bo ho trnh cho mi trng khi b kh. - nh hng ca khng kh: Asp.oryzae l Vi sinh vt hon ton hiu kh, ch pht trin bnh thng khi y oxy. p ng iu kin nui ny, mi trng phi xp, ri thnh lp dy khng qu 2,5-3cm, phng nui phi thong. Theo thc nghim, tho mn cho s h hp ca nm mc Asp.oryzae trong ton b chu k pht trin c cch 1gi, mi trng cn khong 1,7m3 khng kh. Asp. oryzae pht trin bnh thng khi nng CO2 trong khng kh t 8%. - nh hng ca nhit : Nhit nui cng l mt yu t ht sc quan trng i vi s sinh trng ca chng nm mc ny v s to thnh cc enzym amylase. Nhit thch hp cho s pht trin v hnh thnh enzym l 28-320C. Nhit do nm mc ta ra mi trng c th b nng ln 400C hoc hn. Do vy cn gi cho nhit mi trng khng xung di 270C v khng cao hn 360C. Ton b chu k sinh trng ca nm mc cm c th chia lm hai thi k: + Thi k trng v ny mm ca nh bo t trong 11 gi u. Trong thi k ny cn phi cung cp nhit cho phng nui khng hn 23-300C . m tng i ca khng kh l 96-100%. + Thi k sinh trng nhanh ca h si (c th t t 4-18 gi). - Thi gian nui nm mc: Hu ht cc chng nm mc Asp. oryzae c hot ng cc i ca amylase khong gi th 30-38, ri sau l cc i ca protease gi th 36-42. Mt s chng cho hai cc i ca enzym, v d: cc i ca enzym amylase gi th 36 v gi th 60. Do , thi gian nui mc c th ko di t 60-72 gi. Thi gian nui c lng amylase cc ln thng c xc nh bng thc nghim. S to bo t l hin tng khng mong mun v thng lm gim hot
Trang 32
lc ca enzym. i vi Asp. Oryzae, s to hot lc ti a thng kt thc khi nm mc bt u sinh bo t. - nh hng ca pH: pH thch hp cho Asp.oryzae l mi trng acid yu khong 5,5-6.5. Cc mi trng t nhin t cm, nh thng c sn pH khong ny nn khng cn iu chnh. i khi kh nng sinh bo t ca nm mc b yu hoc mt hn. khi phc kh nng ny c th nui nm mc trong nh sng khuch tn trong mt vi th h.
TI LIU THAM KHO

www.en.wikipedia.org www.mrc-lmb.cam.ac.uk www.schmohz.com www.baigiang.bachkim.vn www.elearning.hueuni.edu.vn www.thuocbietduoc.com.vn
Trang 33

Bao Cao Enzyme Amylase Va Cac Ung Dung

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Bao Cao Enzyme Amylase Va Cac Ung Dung

Uploaded by

Copyright:

Available Formats

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

TI LIU THAM KHO .33

I. VAI TR CA ENZYME AMYLASE:

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Enzym - amylase(EC 3.2.1.1) hay 1-4 glucanohydrolase

Enzyme Pullulanase (EC 3.2.1.41 ) hay - dextrin 6 glucosidase.

Enzyme Isoamylase (EC 3.2.1.68) hay glycogen 6 glucanohydrolase.

Enzyme amylase (EC 3.2.1.2).

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Enzyme - amylase (EC 3.2.1.3) hay glucose amylase 3. c Tnh:

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

C Ch Tc Dng: Giai on dextrin ha:

Cc giai on ca qu trnh thy phn tinh bt ca -amylase

Tinh bt Giai on ng ha: Dextrin Amylase Maltose

dextrin phn t lng thp disaccharide&monosaccharide. poliglucose.

tetra v trimaltose oligosacharide maltotriose

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

NEOGADINE elixir 150ml

ng Dng Ca Enzyme Amylase Trong Cng Nghip Dt

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

ng Dng Ca Enzyme Amylase Trong Vic Ty Mu Giy.

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng

GVHD: TS.Nguyn Cng H

Bo Co Enzyme Amylase & Cc ng Dng