Mn

Cng ngh enzyme

GVGD: Th.s H Th Ngn H
Lp: DH11TP
TRNG I HC AN GIANG
Khoa Nng Nghip Ti Nguyn Thin Nhin
B mn: Cng ngh thc phm
1
2
3
4
5
Tng quan v enzyme
Mt s enzyme
Cc yu t nh hng n enzyme
Quy trnh sn xut ru
Quy trnh sn xut bia
Ni dung
Tng quan v enzyme
Protein
Tham gia xc tc
cc phn ng ha
hc trong v ngoi
c th
S dng enzyme trong
sn xut ru bia
-amylase amylase
Enzyme
trong sn
xut
ru bia
Amylase (, , )
Protease
Pullulanase
Cellulase
Mt s enzyme
c im
Enzyme amylase l protein c phn t lng thp, thng nm
trong khong 50.000 n 60.000Dal. Chui mch acid amin Enzyme
amylase u c cu trc bc 3.
Enzyme amylase
Cu trc khng gian ca enzyme -amylase
Tnh
cht
Enzyme amylase
pH ti thch: amylase t nm si l 4.0
4.8
Ngun gc khc nhau bn i vi tc
dng ca acid cng khc khc nhau.
Nhit ti thch ca amylase l 50C
v b v hot 70C
Trong dung dch m pH = 4.7, amylase
ca Asp.orysee rt nhy vi tc ng ca
nhit cao
1
2
3
4
Enzyme amylase
L mt Enzyme ngoi bo (exoenzyme)
Phn gii bt u t u khng kh ca cc nhnh
ngoi cng c cht
Phn ct lin kt l,4glucoside nhng khi gp lin
kt 1,4 glucoside ng k cn lin kt
l,6glucoside th ngng tc dng
c im
Cu trc khng gian ca Enzyme amylase
Tnh
cht
Tm xc tc c cha nhm SH, nhm X-COOH,
vng imidazol
Khng thu phn tinh bt sng, m ch phn ct
tinh bt h ho.
C hot tnh khi khng c Ca
2+
, v b kim hm
bi Cu
2+
, Hg
2+
,...
-amylase hot ng thch hp pH=4.2 5.6
Ngun gc t vi khun th bn nhit hn ca thc
vt v nm mc
Enzyme amylase
c im
C nhiu tn gi
khc nhau:
amyloglucosidas
e,
glucoamylase,...
Thuc nhm
enzyme ngoi
bo exoamylase.
Do vi sinh vt to
ra.
Khi lng phn
t: 27.000 -
112.000 Dal.
Tnh cht
c to nn t
hai isoEnzyme I v
II.
Amyloglucosidas
e I t hp th v
thu phn tinh bt
trng thi rn.
amyloglucosidase
II khng c c hai
tnh cht ny.
Hot tnh
- pH = 3.5 5.5
- Nhit 50C
- Bn vi acid hn
amylase nhng
km bn hn trong
ru, acetone v
khng c bo v
bi Ca
2+
.
Enzyme amylase
c im
Xc tc s thy phn lin kt peptit (CO NH)
trong phn t protein v cc c cht tng t.
C kh nng lin kt este v vn chuyn acid amin.

Enzyme protease
Enzyme protease
Enzyme protease
Cacboxypeptidase
Aminopeptidase
Dipeptidhydrolase
Proteinase
Thy phn lin kt
peptit u nit ca
mch polypeptit
Thy phn lin kt
peptit u cacbon
ca mch polypeptit
Thy phn cc
lin kt dipepit
Thy phn cc lin
kt peptid ni mch
Ngun
gc
T
Vi khun Aerbacter
aerogenes
T
Vi khun Klebsiella
Enzym Pullulanase
Enzyme Pullulanase
Enzym Pullulanase
Thu phn lin kt 1,6 glycoside trong
tinh bt v glycogen
Thu phn nhng dextrin phn t thp
Di tc ng ca iso amylase hay
pullulanase to thnh s l glucose
Hot tnh ti u ca chng pH = 6 v
nhit ti u 60
0
C.
1
2
3
4
Tnh
cht
Enzym Pullulanase
Enzyme cellulase
Thu phn lin kt 1,4--glucoside trong cellulose to ra
sn phm glucose
c im
Phc h
cellulase
Dng 1 l Endo--1,4-glucanase
Dng 2 l Exoglucanase
Dng 3 l -glucosidase
Cu trc khng gian enzyme cellulase
Enzyme cellulase
Enzyme cellulase
Thy phn ngu nhin bn trong phn t
cellulose to ra cc loi oligosaccharide.
Thy phn u kh v u khng kh ca
chui cellulose
Endogluacanase
Xc tc qu trnh ct lin kt -1,4-
glucoside trong cellulose.
Sn phm l cc cellulose phn t nh,
cellobiose v glucose.
Endocellulase
Ct 2 hoc 4 n v glucose t u
khng kh ca chui cellulose.
Exocellulase
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Yu t
nh hng
Nhit
pH
Cht hot ha
Cht kim hm
Nng
enzyme, c cht
Yu t khc
Cc yu t nh hng n enzyme
Nhit

Ti
thch
Trung tm
hot ng
Cao
Thp
Bin tnh
Khng mt
hot tnh m
ch b gim.
Mt i cu hnh chun v
khng ph hp vi c cht
Nhit ti thch
t 40 60
o
C
nh hng ca nhit
nh hng pH
Thay i khng thun nghch
phm vi pH cc hp
Hai sn ca ph ti thch ra s phn
ly nhm prosthetic hay coenzyme
Lm thay i mc ion ha hay
phn ly c cht
Thay i i lc lin kt ca
enzyme vi c cht
nh hng cht hot ha
L cht lm tng kh nng xc tc chuyn ha c cht
thnh sn phm
Thng thng l nhng cation kim loi hay nhng hp
cht hu c nh cc vitamin tan trong nc
Cc cation kim loi c th c tnh c hiu, tnh i
khng v tc dng cn tu thuc vo nng
nh hng cht kim hm
L cht c tc dng lm gim hot hay lm enzyme khng
cn kh nng xc tc bin c cht thnh sn phm.
Phn loi
Thun
nghch
Cnh tranh
Phi cnh
tranh
Khng thun
nghch
nh hng nng enzyme, c cht
Vn tc t
cc i khi
ton b
enzyme lin
kt vi c
cht
Nng
enzyme cng
cao th vn
tc phn
ng enzyme
cng ln
Nng enzyme
C cht
nh hng cc yu t khc
Sng siu m: Tc ng rt khc nhau i vi tng loi
enzyme, c enzyme b mt hot tnh, c enzyme li khng
nh hng.
nh sng: C nh hng khc nhau n tng loi enzyme,
cc bc sng khc nhau c nh hng khc nhau, thng
nh sng trng c tc ng mnh nht, nh sng c tc
ng yu nht. nh sng t ngoi cng c th gy nn
nhng bt li.
S chiu in: in chiu vi cng cng cao th tc
ng ph hy cng mnh
ngui
Trn bnh
men
Ln men Chng ct
Sn
ph
m
Quy trnh sn xut ru go
Nguyn
liu
Ngm nc Nu chn
ng dng cu enzyme trong sn xut ru
Nguyn liu
Gii thch quy trnh
Go, np hoc cc loi nng sn c hm lng tinh bt cao
thng cha khong 14% nc v trn 74% tinh bt.
Nguyn liu tt khng su mt, mc, khng ln tp cht khc,

Cho nguyn liu ngm trong nc nhm loi b tp cht
bn...
Trong qu trnh ngm nguyn liu ht nc v tr nn
mm hn, rt ngn thi gian nu.
Ngm nc
Nu chn
Vai tr ca amylase: amylase hot ng mnh trong giai
on u ca qu trnh ng ha.
Nguyn nhn: C s thay i pH trong thi gan ng ha v
gii hn hot ng ca a-amylase ti ng khng vt qu 70
80%.
=> Vai tr enzyme ny l lm dch ha nhanh giai on
nu v c giai on u ca qu trnh ng ha, dextrin
v tch t ng.
Gii thch quy trnh
Glucoamylase thy phn kin kt 1,4 glucose trong cc
polysacharide.

Ngoi ra enzyme ny cn c kh nng phn ct lin kt
1,6 glucoside.

Phn ln cc glucoamylase hot ng pH khng cao.

Nhit ti u l 55 60C.

Sn phm cui cng ca qu trnh thy phn ny l glucose
Nu chn
Gii thch quy trnh
Vai tr ca glucoamylase
Nu chn
Gii thch quy trnh
Vai tr ca pululanase
Tham gia thy phn lin kt 1,6 glucoside trong cc poly v
oligosacharide
Phn ct lin kt 1,6 glucosidetrong trng hp lin kt ny
c bao quanh tt c cc pha nh lin kt 1,4 glucoside
Dextrin c phn t thp, d b pululanse phn hy to thnh t 2
gc maltose, ni vi nhau bng lin kt 1,6 glucoside
Nu c s kt hp ca amylase v pululanase, dextrin s b
thy phn hon ton
Nu chn
Gii thch quy trnh
Vai tr ca h enzyme cellulase
Tham gia phn hu cc polysacharide khng phi l glucide
B sung enzyme trn vo trong qu trnh ng ho hm
lng ng galactose, arabinose, cylose, glucose tng ln
H enzyme cellulase c coi nh bin php tng cng kh
nng ng ho v lm tng hiu sut thu cn sau ln men
Nu chn
Gii thch quy trnh
Vai tr ca enzyme protease
Khi x l nguyn liu bng ch phm protease, trong
dung dch ng ha tch t nhiu amino acid
Amino acid l thnh phn rt cn thit cho s pht
trin ca nm men
Ln men m
Ln men
L qu trnh to iu kin cho enzym amylase ca nm mc,
vi khun xc tc thy phn tinh bt.
Gii thch quy trnh
Cm trn men c trong 5 10 gi mc mc u c khi
cm, sau vun thnh ng, ph kn bng vi v gi ni
thong mt (nhit 28 32
o
C) trong 2 3 ngy
Mc ch: To iu kin cho nm mc v vi khun pht trin, tit
ra enzym ng ha tinh bt
L qu trnh nm men s dng ng to ra ln men
ru.
Thi gian khong 2 3 ngy.
Cm ru trong iu kin kn, nm mc ngng pht trin
nhng enzyme to thnh vn tip tc thy phn tinh bt.
Trong giai on ny, nm men pht trin mnh nh lng
oxi ha tan trong nc v lng ng to thnh, sau
chuyn sang ln men ru.
Ln men lng
Ln men
Gii thch quy trnh
Chng ct
Gii thch quy trnh
Nhm tch hn hp ru v nc c nhit si khc nhau.
Khi chng ct, ru c tch ra khi nc nh nhit
bc hi thp hn nc
Tin hnh bng cch un si hn hp ln men, hi bay ln
c dn qua ng dn v c lm lnh bng cch cho ng
dn i qua bn nc ngng t ru bn trong lng ng
Dung dch ru thu c trong sut, c mi thm c trng
v nng ru s gim dn theo thi gian chng ct
Qu trnh ln men kt thc th tin hnh chng ct thu
c ru thnh phm

Ln men
ph
Lm
trong bia
Bo ho
CO
2

Chit rt
chai
Thanh
trng
Thnh
phm
i mch Ngm
m
mm
Sy Nghin
ng
ho
Lc b
Nu
Lc b
malt
Lm lnh
Ln men
chnh
Quy trnh sn xut bia
ng dng cu enzyme trong sn xut bia
Cc enzym trong giai on m mm
Enzym sitase
Enzym protease
Enzym amylase
Enzym lipase
Enzyme
amylophotphatase
Gii thch quy trnh
Tc ng ca Sitase (glucanasepentosanase)
Thnh
hemicellulose
ng n pentose,
hexose v cc s
phm khc
C ch hot ng
Enzym sitase bt u hot ng ni nh.
u tin u tin l s ha lng mng ca t bo gn
k vi ng.
Enzym tc ng theo trc ca ht.
Kh nng hot ng ca enzym sitase quyt nh kh
nng ny mm
Cc yu t nh hng n hot ng ca enzym sitase

Nhit
Bn cht ca ht i mch
Gii thch quy trnh
Hot ng ca protease
Protease c tit ra t phi ht v hot ng ngay
khi bt u m mm.
Tc ng trc tin vo cc t bo ni nh.
protein
Cc acid amin ho tan
trong dch thc
Chui
peptid
Abumo,
pepton
Gii thch quy trnh
Hot ng ca enzyme amylase

Cc amylase tn cng v n mn b mt ht tinh
bt.

Cc t bo gn mm b tn cng mnh hn cn
cc t bo khc hu nh khng b tn cng.

Maltose c to thnh di dng ho tan v di
chuyn n mm.
Gii thch quy trnh
Hot ng ca enzym lipase
Thu phn lipid
Gii thch quy trnh
Hot ng ca enzyme amylophotphatase
Enzyme phytase thu phn cc
este photphoric

Gii thch quy trnh
Qu trnh ng ho
Thu phn
tinh bt
Thu phn
-glucan
Thu phn
protein
Qu trnh h ho
Qu trnh dch ho
Qu trnh ng ho
Gii thch quy trnh
Qu trnh dch ho
Cc gc glucose
(amylose v
amylopectin)
-amylase
-amylase

Thu phn
hnh thnh
mch ngn
hn.
Gim nht
Gii thch quy trnh
Qu trnh ng ho
Amylose v
amylopectin
-amylase
Thu phn
thnh dextrin
cha 7 12
gc glucose
Dextrin
-amylase
Phn ct
thnh hai gc
maltose u
khng kh
Ngoi ra, amylase tham gia thu phn polysaccharide t
u khng kh tun t tng gc glucose. Ct loeen kt
1,4; 1,6; 1,2 v 1,3 glucozid. Sn phm to thnh l
glucose v oligosaccharide.
Gii thch quy trnh
Thu phn -glucan
Gii thch quy trnh
Thnh t bo ni nh: cha mt mng li cc protein,
cellulose v hemicellulase vng chc, cc cht ny lin kt vi
nhau bi glucan
glucan lm tng nht ca dch ng v bia gy
cn tr qu trnh lc
Thu phn protein
Thu phn protein nh enzym protease.
Protein b phn ct dn thnh cc mch nh hn v to
thnh cc acid amin.
Cc hp cht cao phn t to thnh trong qu trnh
thy phn protein li tip tc b phn ct cho nn thi
gian to thnh chng khng cn ko di, c bit l
cc cht keo, to bt b phn hy cng nhit .
Gii thch quy trnh
Lc b
Tch dch ng hay dch hm vi v v nhng phn
ni nh ca ht khng tan. Ngoi ra, loi b nhng cht
khng mong mun nh: kim loi nng, tanin,
Nu vi hoa houblon
n nh thnh phn v to cho bia c mi thm v v
ng c trng ca hoa houblon.
Gii thch quy trnh
Loi b cn, cht huyn ph v hoa houblon. To iu
kin thun li cho vi sinh vt hot ng
Lc trong
Lm lnh
To nhit thch hp cho vi sinh vt ln men hot
ng.
Gii thch quy trnh
Ln men chnh
Chuyn ho cc cht ho tan trong dch ng thnh
C
2
H
5
OH, CO
2
v cc sn phm ph khc (sinh tng
hp trong qu trnh hot ng sng ca t bo nm
men) to thnh bia theo yu cu k thut v cht lng
thnh phm.
Gii thch quy trnh
Ln men ph
Tip tc ln men phn cht kh cn st li sau ln
men chnh, bo ha CO
2
v tng cng mi v cho
bia, thc hin qu trnh chn ca bia (n nh
cht lng bia), lm trong bia; a bia v nhit
thp hn ch s xm nhp v ph hy ca vi
sinh vt.
Gii thch quy trnh
Li
tm
Lm trong bia
Tch cn l lng, tch cc t bo nm men sng v
xc t bo non
Tng gi tr cm quan, n nh thnh phn c hc,
lm tng bn sinh hc, bn keo ca bia
Phng
php
Lc
Lng
Gii thch quy trnh
Bo ha CO
2

CO
2
b sung vo phi m bo khng cha O
2
v
vi sinh vt tp nhim
Cc cng on lm vic nhit cao v p sut
thp lm hm lng CO
2
gim
CO
2
c b sung vo bia nh cc ng ng dn
di kt hp vi cc on ni m ha tan vo bia.
Gii thch quy trnh
Chit chai v thanh trng
Bia sau khi bo ha CO
2
phi c chit chai ngay
Khng tn tht bia
Rt lng
Khng nh hng cht lng
Yu
cu
Gii thch quy trnh
Chit chai v thanh trng
Vi sinh vt gy h hng bia c th xm nhp vo sn
phm do s thiu v sinh
Phng php
thanh trng
Thanh trng
nhanh (nng
nhit n 68
72
o
C trong 50
giy, sau
lm lnh)
Chit bia nng (chai sau khi
ra khng cn lm ngui)
Thanh trng
tunnel (nhit
60
o
C trong
20 pht)
Gii thch quy trnh