L M U

Ngy nay, cng vi su pht trin manh m

ca Cng ngh sinh hoc, cc ch phm
enzyme duoc sn xut cng nhiu v duoc
s dung trong hu ht trong cc lnh vuc
nhu: ch bin thuc phm, nng nghip,
chn nui, y t. Hng nm luong enzyme
duoc sn xut trn th gii dat khong trn
300.000 tn vi gi trj trn 500 triu USD,
duoc phn phi trong cc lnh vuc khc
nhau [4].
Phn ln enzyme duoc sn xut quy m
cng nghip du thuc loai enzyme don
cu t, xc tc cho phn ng phn hy.
Khong 75% ch phm l enzyme thy
phn duoc s dung cho vic thy phn co
cht tu nhin [4].
Protease l enzyme duoc s dung nhiu
nht hin nay trong mt s ngnh sn xut
nhu: ch bin thuc phm (dng tu sa lm
pho mt, lm mm thjt, b sung d lm
tng cht luong sn phm trong sn xut
bia, x l ph phu phm trong ch bin
thuc phm.), sn xut cht ty ra, thuc
da, y t, nng nghip.
Qua nhiu nm, vic gia tng s dung vi
sinh vt nhu l mt ngun cung cp
protease d ci thin dng k hiu qu sn
xut v sn phm duoc tao ra nhiu hon.
Tuy nhin gi thnh ch phm protease
cn kh cao, do d cng han ch vic s
dung rng ri enzyme trong sn xut. Cc
ch phm thu duoc sau qu trnh nui cy
sn xut enzyme chua phi l ch phm c
d tinh khit cao v protein chi chim 20-
30% [6]. V vy, vic nghin cu ci tin
phuong php tch v tinh ch enzyme
nhm thu duoc ch phm c d tinh khit
cao rt cn thit. tch v tinh ch
enzyme ni ring v protein ni chung
thung c mt loat phuong php ha-l v
ha hoc khc nhau. C th chia lm ba
nhm phuong php sau:
- Phuong php kt ta
- Phuong php sc k
- Phuong php phn tch h hai pha nuc
Protease phn b thuc vt, dng vt, vi
sinh vt. Tuy nhin ngun enzyme vi sinh
vt phong ph nht, c hu ht cc vi
sinh vt nhu vi khun, nm mc v xa
khun. C th ni vi sinh vt l ngun
nguyn liu thch hop nht d sn xut
enzyme quy m ln dng trong cng
ngh v di sng.
CHUONG 1. TNG QUAN T LEU
1.1. Tnh hnh nghin cu enzyme.
1.1.1. Vn d nghin cu enzyme nuc
ta
Hu nhu moi phn ng ho hoc trong co
th sng du cn phi c vai tr xc tc
ca enzyme - cht xc tc sinh hoc. Chnh
v vy, cc nghin cu v enzyme d thu
ht su quan tm ca cc cn b ho sinh
hoc, sinh hoc thuc nghim v nhiu nh
nghin cu cc lnh vuc lin quan khc.
Cc nghin cu nhm theo hung tch,
tinh sach enzyme, tao cc ch phm c d
sach khc nhau, nghin cu cu trc, mi
lin quan gia cu trc v hoat tnh sinh
hoc ca enzyme, kh nng ng dung
enzyme trong thuc t. Nghin cu v cng
ngh enzyme d duoc tin hnh bi nhiu
tc gi nhu s dung ph tang ca l m d
sn xut pancrease, pepsin, trypsin... s
dung mm ma d sn xut amylase. c
nhng th nghim cng ngh nhu sn xut
amino acid t nhng tm bng protease,
bt protein thjt bng bromelain t dot da,
ln men ruou bng enzyme c djnh trn
ct. Cng d c nhng nghin cu s dung
peroxydase, cyt-P450 trong ch tao
biosensor v thuc pht hin cht dc...
Trong lnh vuc y duoc, vic nghin cu su
v co ch tc dung ca mt s enzyme
nhm muc dch kin tao nn mt s thuc
dng diu trj mt s bnh dc bit l tao ra
mt s ch phm thuc chng suy dinh
dung tr em, dng thi d tin hnh sn
xut dai tr. y l mt dng gp rt thit
thuc v kjp thi trong vic phng chng suy
dinh dung nuc ta.
1.1.2. Cng ngh sn xut enzyme
Protease trn th gii.
Trong nhng nm gn dy, gi trj thuong
mai ca cc enzyme cng nghip trn ton
th gii dat khong 1 t USD, trong d ch
yu l cc enzyme thy phn (75%), v
protease l mt trong ba nhm enzyme ln
nht s dung trong cng nghip (60%) [4].
1.2. Tng quan v enzyme Protease.
1.2.1. Gii thiu chung.
Nhm enzyme protease (peptit hidrolase
3.4) xc tc qu trnh thu phn lin kt lin
kt peptit (-CO-NH-)n trong phn t protein,
polypeptit dn sn phm cui cng l cc
axit amin. Ngoi ra, nhiu protease cng c
kh nng thu phn lin kt este v vn
chuyn axit amin.
H2N- CH- CO- NH- CH- CO- .- NH- CH-
COOH
R1 R2 RX
+H2O
H2N- CH- COOH + H2N- CH- CO . NH-
CH- COOH
R1 R2 RX

Hnh 1.1. M hnh enzyme Protease thy
phn phn t Protein.
Protease cn thit cho cc sinh vt sng,
rt da dang v chc nng t mc d t
bo, co quan dn co th nn duoc phn b
rt rng ri trn nhiu di tuong t vi sinh
vt (vi khun, nm v virus) dn thuc vt
(du d, da...) v dng vt (gan, da dy
b...). So vi protease dng vt v thuc
vt, protease vi sinh vt c nhng dc dim
khc bit. Truc ht h protease vi sinh vt
l mt h thng rt phc tap bao gm nhiu
enzyme rt ging nhau v cu trc, khi
luong v hnh dang phn t nn rt kh
tch ra dui dang tinh th dng nht.
Cng do l phc h gm nhiu enzyme
khc nhau nn protease vi sinh vt thung
c tnh dc hiu rng ri cho sn phm thu
phn trit d v da dang.

Hnh 1.2. Cu trc khng gian enzyme
Protease.
1.2.2. Phn loai Protease
Protease (peptidase) thuc phn lp 4 ca
lp th 3 (E.C.3.4) .
So d phn loai duoc tm tt trn hnh 1.1:
Protease duoc phn chia thnh hai loai:
endopeptidase v exopeptidase.
* Dua vo vj tr tc dng trn mach
polypeptide, exopeptidase duoc phn chia
thnh hai loai:
+ Aminopeptidase: xc tc thy phn lin
kt peptide du N tu do ca chui
polypeptide d gii phng ra mt amino
acid, mt dipeptide hoc mt tripeptide.
+ Carboxypeptidase: xc tc thy phn lin
kt peptide du C ca chui polypeptide
v gii phng ra mt amino acid hoc mt
dipeptide.
* Dua vo dng hoc ca co ch xc tc,
endopeptidase duoc chia thnh bn nhm:
+ Serin proteinase: l nhng proteinase
cha nhm OH ca gc serine trong trung
tm hoat dng v c vai tr dc bit quan
trong di vi hoat dng xc tc ca
enzyme. Nhm ny bao gm hai nhm nh:
chymotrypsin v subtilisin. Nhm
chymotrypsin bao gm cc enzyme dng
vt nhu chymotrypsin, trypsin, elastase.
Nhm subtilisin bao gm hai loai enzyme vi
khun nhu subtilisin Carlsberg, subtilisin
BPN. Cc serine proteinase thung hoat
dng manh vng kim tnh v th hin
tnh dc hiu co cht tuong di rng.
+ Cysteine proteinase: Cc proteinase
cha nhm SH trong trung tm hoat dng.
Cystein proteinase bao gm cc proteinase
thuc vt nhu papayin, bromelin, mt vi
protein dng vt v proteinase k sinh
trng. Cc cystein proteinase thung hoat
dng vng pH trung tnh, c tnh dc hiu
co cht rng.
+ Aspartic proteinase: Hu ht cc aspartic
proteinase thuc nhm pepsin. Nhm
pepsin bao gm cc enzyme tiu ha nhu:
pepsin, chymosin, cathepsin, renin. Cc
aspartic proteinase c cha nhm carboxyl
trong trung tm hoat dng v thung hoat
dng manh pH trung tnh.
+ Metallo proteinase: Metallo proteinase l
nhm proteinase duoc tm thy vi khun,
nm mc cng nhu cc vi sinh vt bc cao
hon. Cc metallo proteinase thung hoat
dng vng pH trung tnh v hoat d gim
manh dui tc dung ca EDTA.
Ngoi ra, protease duoc phn loai mt
cch don gin hon thnh ba nhm:
- Protease acid : pH 2-4
- Protease trung tnh : pH 7-8
- Protease kim : pH 9-11
CHUONG . TUONG V PHUONG
PHP NGHN CU.
2.1.i tuong nghin cu.
Vi sinh vt: ch yu vi khun, nm mc, xa
khun.
Vi khun
Luong protease sn xut t vi khun duoc
uc tnh vo khong 500 tn, chim 59%
luong enzyme duoc s dung [6].
Protease ca dng vt hay thuc vt chi
cha mt trong hai loai endopeptidase
hoc exopeptidase, ring vi khun c kh
nng sinh ra c hai loai trn, do d
protease ca vi khun c tnh dc hiu co
cht cao. Chng c kh nng phn hy ti
80% cc lin kt peptide trong phn t
protein [6].
Trong cc chng vi khun c kh nng tng
hop manh protease l Bacillus subtilis, B.
mesentericus, B. thermorpoteoliticus v mt
s ging thuc chi Clostridium. Trong d, B.
subtilis c kh nng tng hop protease
manh nht (Nguyn Trong Cn v cs,
1998). Cc vi khun thung tng hop cc
protease hoat dng thch hop vng pH
trung tnh v kim yu.
Cc protease trung tnh ca vi khun hoat
dng khong pH hep (pH 5-8) v c kh
nng chju nhit thp. Cc protease trung
tnh tao ra djch thy phn protein thuc
phm t dng hon so vi protease dng vt
v tng gi trj dinh dung. Cc protease
trung tnh c kh nng i luc cao di vi
cc amino acid ua bo v thom. Chng
duoc sinh ra nhiu bi B. subtilis, B.
mesentericus, B. thermorpoteoliticus v mt
s ging thuc chi Clostridium [2].
Protease ca Bacillus ua kim c dim
dng din bng 11, khi luong phn t t
20.000-30.000. n djnh trong khong pH 6-
12 v hoat dng trong khong pH rng 7-12
.
Nm
Nhiu loai nm mc c kh nng tng hop
mt luong ln protease duoc ng dung
trong cng nghip thuc phm l cc chng:
Aspergillus oryzae, A. terricola, A.
fumigatus, A. saitoi, Penicillium
chysogenum). Cc loai nm mc ny c
kh nng tng hop c ba loai protease:
acid, kim v trung tnh. Nm mc den tng
hop ch yu cc protease acid, c kh
nng thy phn protein pH 2,5-3 [2].
Mt s nm mc khc nhu: A. candidatus,
P. cameberti, P. roqueforti. cng c kh
nng tng hop protease c kh nng dng
tu sa s dung trong sn xut pho mt.
Xa khun
V phuong din tng hop protease, xa
khun duoc nghin cu t hon vi khun v
nm mc. Tuy nhin, ngui ta cng d tm
duoc mt s chng c kh nng tng hop
protease cao nhu: Streptomyces grieus, S.
fradiae, S. trerimosus. [2].
Cc ch phm protease t xa khun duoc
bit nhiu l pronase (Nht) duoc tch
chit t S. grieus, enzyme ny c dc tnh
dc hiu rng, c kh nng thy phn ti
90% lin kt peptide ca nhiu protein
thnh amino acid. Lin X (c), ngui ta
cng tch duoc ch phm tuong tu t S.
grieus c tn l protelin.
T S. fradiae cng c th tch chit duoc
keratinase thy phn karetin. M, ch
phm duoc sn xut c tn l M-Zim dng
trong sn xut da [2]. Protease t S. fradiae
cng c hoat tnh elastase cao, do d
chng duoc dng trong cng nghip ch
bin thjt.
Hu ht cc protease phn ct protein
cc lin kt dc hiu, v th c th s dung
cc enzyme ny theo chiu phn ng tng
hop d tng hop cc lin kt peptide djnh
truc. Yu t tng cung qu trnh tng
hop bao gm pH, cc nhm carboxyl hoc
nhm amine duoc lua chon d bo v, kh
nng kt ta sn phm, phn ng trong h
hai pha lng.
2.2. Phuong php nghin cu.
Qu trnh sn xut cc ch phm enzyme
vi sinh vt bao gm cc giai doan ch yu:
2.2.1. Tuyn chon v ci tao ging vi sinh
vt cho enzyme c hoat luc cao.
chon ging vi sinh vt c kh nng sinh
tng hop enzyme cao, ngui ta cth phn
lp t mi trung tu nhin hoc c th
dng cc tc nhn gy dt bin tc dng
ln b my di truyn hoc lm thay di dc
tnh di truyn d tao thnh cc bin chng
c kh nng tng hop dc bit hu hiu
mt loai enzyme no d, cao hon hn
chng gc ban du.
2.2.1.1. Phuong php gy dt bin.
y l phuong php hay duoc dng nht
nhm d:
-Tao nhng dt bin bj gim kh nng sinh
tng hop repressor hoc tng hop
repressor c i luc thp vi gene opertor.
-Tao nhng dt bin tng hop enzyme c
cu trc bc 1 thay di do d c th gim
d thay di vi kiu km hm theo co ch
lin h nguoc.
Nu su thay di cu trc bc 1 xy ra
vng trung tm hoat dng hoc gn d
th c th lm thay di r rt hoat tnh ca
enzyme.
-Gy dt bin doan gene hoat ha
promotor d lm tng p luc ca n di vi
ARN-polymerase do d lm tng tc d sao
chp m.Dng bin php ny c th lm
tng luong glucoza-6-
phosphatdehydrogenaza ln 6 ln.
Hin tuong dt bin thung lin h vi su
thay di mt gene, chng han bj "li mt
bazo khi ti tao phn t ADN.[1]
tao mt dt bin gene c th dng tc
nhn vt l (tia t ngoai, tia phng xa) hay
ha hoc (cc ha cht) tc dung ln t bo
sinh vt.
2.2.1.2. Phuong php bin nap.[1]
L su bin di tnh trang di truyn ca mt
ni vi sinh vt dui nh hung ca ADN
trong djch chit nhn nhn duoc t t bo
ca vi sinh vt khc. dy yu t bin nap
l AND. Su chuyn vt liu di truyn (ADN)
t t bo cho dn t bo nhn c th xy ra
trong ng nghim (invitro) khi cho t bo
nhn tip xc vi djch chit t t bo cho
m khng c su tip xc gia cc t bo.
Cc t bo c th nhn bt ky loai ADN no
ch khng di hi phi l ADN t cc ging
ho hng. Tuy nhin t bo chi c th nhn
mt s doan ADN nht djnh, thung khng
qu 10 doan. Cc doan ADN duoc di
truyn trong bin nap c M=106-107 v
phi c cu trc xon kp. Hin tuong bin
nap ph bin nhiu loi vi khun nhu:
Diplococus, Staphylocus, Hemophilus,
Agrobacterium, Rhizobium, Bacillus,
Xantomonas.
2.2.1.3. Phuong php tip hop gene.[1]
Khc vi bin nap, dy vt liu di truyn
chi duoc truyn t t bo cho dn t bo
nhn khi hai t bo tip xc vi nhau. Do
vy cc vi sinh vt c kh nng bin nap th
s khng c kh nng tham gia tip hop
gene na. Hin nay qu trnh tip hop gene
d duoc nghin cu mt s loi vi khun
nhu E.coli, salmonella, Pseudomonas
aeruginosa.
2.2.1.4. Phuong php ti nap[1]
Vt liu di truyn (ADN) duoc chuyn t t
bo cho sang t bo nhn nh vai tr trung
gian ca thuc khun th (phage). Trong
qu trnh ti nap, cc doan AND duoc
chuyn t t bo cho dn t bo tip hop
vi AND ca t bo nhn. Do d lm bin
di tnh cht di truyn ca t bo nhn.
2.2.2. Phuong php bo qun ging vi sinh
vt.
Khi s dung vi sinh vt d sn xut enzyme
cn chon ging thun chng, d duoc kim
tra dy d v cc dc tnh ha sinh, vi sinh,
nui cy v cn dc bit luu dn diu kin
bo qun ging. Thuc t khi bo qun
ging gc trong mt thi gian c th tao ra
cc bin dj ngu nhin khng mong mun
do d djnh ky phi cy chuyn v kim tra
lai cc dc tnh ban du.
Phuong php cy chuyn.
y l phuong php ph bin nht d thuc
hin bng cch gi ging trn mi trung
thach (thach nghing, hp petri,.) vi
thnh phn mi trung nui cy v diu
kin nui cy thch hop cho ging vi sinh
vt d. Sau khi ging d moc tt cn bo
qun nhit d lanh 3-40C v sau mi
tun phi cy chuyn lai. Khi cy chuyn
chi ly bo t hoc khun lac m khng
nn ly c mi trung dinh dung d dm
bo khng chuyn cc sn phm trao di
cht vo mi trung mi (c th gy bin
di bt loi khng th lung ht duoc). Nu
l xa khun th khng nn bo qun ging
trn mi trung thach m nn gi trong dt
d kh trng.
ko di thi gian bo qun ging t
hng thng dn 1 nm, ngui ta ph 1 lp
paraphin lng d tit trng trn b mt
ging d han ch su pht trin ca n. Cn
luu chi ph lp du sau khi cy vi sinh vt
dat dn d chn sinh l.
Phuong php cy chuyn rt c hiu qu
d bo qun cc ging nm men, vi khun
v rt hu hiu, d dng trin khai ging ra
sn xut ln, han ch cc tai bin c th
dn dn hu hng ging gc.
Phuong php lm kh
Bng cch gi ging trn ct, dt, silicagen
trong diu kin kh ro (tt c du duoc
kh trng cn thn). Trong diu kin nhu
vy s han ch su pht trin tip tuc ca
ging khi bo qun. Phuog php ny rt
hay duoc s dung d bo qun nm mc,
xa khun, mt vi loai nm men, vi khun
thi gian giu ging c th duoc 1 nm.
Phuong php lm kh cng thuc hin don
gin, khng cn dung cu dt tin. Tuy nhin
ging nhu phuong php cy chuyn thi
gian bo qun tuong di ngn.
Phuong php dng kh
ng kh l qu trnh m nuc duoc ly ra
khi mu khi cc mu dang trang thi
lanh su. dy vi sinh vt duoc huyn ph
trong mi trung thch hop v duoc lm
lanh trong mi trung chn khng. Thit bj
dng kh s ht nuc v cui cng mu
duoc lm kh dn mc nht djnh. Mu
duoc hn kn d cho mi trung cha mu
l chn khng. y l phuong php ph
bin c hiu qu cao cho bo qun cc di
tuong vi sinh vt khc nhau nhu nm soi,
nm men, vi khun v mt s virut. Tuy
nhin, phuong php ny t duoc ng dung
di vi to, dng vt nguyn sinh v t bo
dng vt.
Phuong php bo qun lanh su:
i vi phuong php bo qun lanh su th
vi sinh vt duoc bo qun trong mi trung
djch th v nuc cn cho hoat dng sng
ca vi sinh vt bj bt hoat nhit d lanh
su (-196C -> -80 C).
Vi phuong php ny, t bo c th bj v
trong qu trnh lm lanh v lm tan mu.
Mt nguyn nhn dn dn lm v t bo l
vic tch lu cc cht din gii trong mu
bo qun v hnh thnh cc tinh th nuc
trong t bo. khc phuc nhuoc dim
ny ngui ta d b sung cc cht lm han
ch tc d lanh su v lm tan nhanh nhu
glycerol, DMSO (dimethyl sulfoxide). Vic
bo qun theo phuong php lanh su ny
duoc thuc hin cc thang nhit d khc
nhau nhu -20 C, -30 C, -40 C, -70 C, -
140C v -196 C. Ni chung mc nhit d
cao hon -30 C cho hiu qu thp do t bo
chju nng d mui cao sinh ra t cc cht
din gii. Phuong php bo qun ny c
hiu qu vi nhiu nhm vi sinh vt khc
nhau nhu nm soi, nm men, vi khun, xa
khun v virut.
c bit vi phuong php bo qun lanh
su trong nito lng l phuong php van
nng hon c. Phuong php ny thch hop
vi nhiu di tuong vi sinh vt khc nhau
nhu vi khun, nm soi, nm men, virut, to
v c cc dng t bo dng vt. Tuy nhin,
phuong php ny cng bc l mt s
nhuoc dim nhu du tu kinh ph cho thit
bj v din, nito lng hoc ri ro nhu chy
n... c bit phuong php ny khng
thch hop vi cc chng vi sinh vt thung
xuyn dng dn. Ni chung phuong php
ny thung duoc dng vi cc chng vi
sinh vt c nhng dc tnh qu m khng
thch hop vi phuong php dng kh.
2.2.3. Mi trung nui cy vi sinh vt tng
hop enzyme Protease.
Cn phi chon mi trung v thnh phn
mi trung dinh dung c nh hung truc
tip dn su sinh trung v tng hop
enzyme ca vi sinh vt. Trong thnh phn
mi trung phi c d cc cht dm bo
duoc su sinh trung bnh thung ca vi
sinh vt v tng hop enzyme.
c bit luu l d tng su tng hop
enzyme ngui ta thung dua vo hin
tuong cm ng. V nu nhu trong thnh
phn mi trung c cc cht cm ng th
cht d hay sn phm phn gii ca n s
km hm hoc lm yu tc dung km to
ca cht km hm nhm bo dm kh nng
sinh tng hop enzyme d cho khng bj cn
tr. Cht cm ng tng hop enzyme cho
thm vo mi trung nui thung l co
cht tuong ng ca enzyme cn tng hop.
Thnh phn chnh ca mi trung: C, N, H,
O. Ngoi ra cc cht v co: Mn, Ca, P, S,
Fe, K v cc cht vi luong khc.
2.2.3.1. Ngun cacbon.
Thung l hop cht hu co trong d ch
yu l gluxit, ty thuc vo dc tnh ca
enzyme v ni vi sinh vt m ngui ta lua
chon cho thch hop.
C nhiu cht hydratcacbon v cc hop
cht khc l ngun cacbon thch hop di
vi nm mc sinh ra enzyme protease c
hoat luc cao. Cc ngun cacbon c tc
dung dn sinh tng hop proteinase ca
nm mc c th theo th tu:
i vi Asp. flavus 74: fructoza
glucoza sacaroza ramnoza
manoza galactoza arabinora lactoza.
i vi Asp. awamori 200: fructoza
manit sacaroza arabinoza manoza
galactoza lactoza.
i vi Asp. oryzae 79: fructoza
sacaroza maltoza glucoza manit
arabinoza galactoza.
Tinh bt l ngun cacbon ca nhiu chng
vi khun sinh tng hop enzyme Protease.
V du: Vi khun Bac. Subtilis c kh nng
sinh tng hop Protease mi trung tinh
bt >8%.
Nhiu xa khun ua nhit, trong d
Micromonospora vulgaris 42, moc tt v
sinh tng hop protease cao mi trung
c tinh bt. Tng nng d tinh bt t 0,25-
1,5% sinh khi cng tng dng thi vi hiu
xut tng hop enzyme.Nu tng nng d
tinh bt hon na s khng thu duoc kt
qu duong tnh. T s gia cacbon v nito
cn phi l 4: 1. Trn mi trung c
maltoza (0,5- 2%) vi sinh vt pht trin bnh
thung, nhung tng hop protease ngoai
bo bj c ch. Xa khun khng pht trin
duoc trn mi trung c cc ngun
cacbon duy nht l sacaroza, lactoza hoc
arabinoza. Glucoza, manoza, fructoza,
xyloza c trong mi trung (0,2- 5%) lm
c ch sinh trung ca xa khun v sinh
tng hop enzyme.
Ngoi ra mt s loai hydrocacbon cng c
ngun cacbon c 125 chng vi sinh vt.
Chng han chng Ps. Seruginosa c th
dng ho hydrocacbon v c kh nng sinh
tng hop protease c hoat luc cao trn mi
trung n- parafin vi 12, 14 v 16 nguyn
t cacbon, du nng hoc propylenglycol.
Chng khng chi dng ho duoc c
hydrocacbon bo m cn dng ho c
hydrocacbon thom.[1]
2.2.3.2. Ngun nito.
Ngun nito s dung rt phong ph, bao
gm 2 nhm: v co v hu co.
i vi mt s loi nm mc thuc ho (A.
oryzae, A. awamori, A. niger, A. flavus) trn
mi trung c cc ngun nito hu co sinh
tng hop protease axit cao. Trn mi
trung kzapek NaNO3 duoc thay bng c
cazein hoat luc protease ca nm mc
duoc tng ln 3,5 ln, cn khi thay tinh bt
bng glucoza v b sung pepton hoat luc
enzyme tng 6 ln. Nhiu kt qu nghin
cu cho thy sinh tng hop enzyme duoc
nng cao khi trong mi trung c dng thi
c ngun nito hu co v nito v co. Cho
thm vo mi trung c cm m, bt du
tuong d tch cht bo, cc ngun nito v
hu co hoat luc enzyme protease tng 22-
74%. Cn trung hop dng cc ngun nito
v co duy nht trong mi trung s dn
dn ngng sinh tng hop protease ni
chung.
Trong qu trnh nui cy vi khun, trong d
c B. subtilis v B. mesentericus, cc hop
cht nito v co v hu co duoc dng phi
hop trong mi trung s tao diu kin
thun loi cho sinh tng hop protease. Trong
s cc ngun nito v co th NH4, H2PO4 l
tt hon c. Nhng mui khc NH4Cl,
(NH4)2SO4, NH4NO3, NaNO3, KNO3,
Ca(NO3)2 lm gim hoat luc protease ti
30- 50% , cn amylaza gim 7- 10 ln. Cn
trong mi trung chi c ngun nito hu co
hoat luc protease v amylaza cng thp
hon trong mi trung di chng
(NH4)2HPO4 v nuc chit du tuong.
i vi xa khun ua nhit Actynomyces
Vulgaris U2 th pepton l cht cm ng d
sinh tng hop enzyme protease l tt nht.
Cc axit amin c nh hung r rt ti sinh
tng hop enzyme bng vi sinh vt. Glyxin,
alanin, metionin v loxin c tc dung lm
tng hoat luc protease ca chng dt bin
A. oryzae 251- 90 dn 6- 9% v nguyn
chng A. oryzae 132- 63 ti 7- 24%. Nhiu
axit amin c tc dung c ch dn sinh tng
hop enzyme, Valin, axit glutamic, izoloxin
v valin c ch tng hop enzyme B.
megaterium 60%. Cn di vi B. subtilis
cc axit amin c ch trong qu trnh ny l
glyxin, metionin, axit glutamic, alanin,
loxin,.
Ngoi ra, cc bazo purin nhu A (adenin), G
(guanin) v cc dn xut ca chng, ARN
v cc sn phm thu phn cng lm tng
dng k sinh tng hop proteinza vi sinh
vt.[1]
2.2.3.3. Ngun cc nguyn t khong v
cc yu t (cht) kch thch sinh trung
Mui khong rt cn thit cho hoat dng vi
sinh vt.
on Mg2+ c tc dung sinh tng hop v n
djnh cc enzyme c hoat tnh nhit d
cao.
C nhiu yu t nh hung di vi mi
trung nui cy. Nhit d nui cy thng
thung t 25 - 300C. Trj s pH ban du
ca mi trung (ch yu mi trung
nuc) cng c th gy nh hung no d
dn su tao thnh enzyme, nhung khi d
cng cn tnh dn kh nng bin di nhanh
chng chi s d bi vi sinh vt.
thng kh cng rt cn thit cho vic
sinh tng hop enzyme. V vy mi trung
b mt ngui ta thung thm cht xp nhu
tru vo, cn mi trung b su (mi
trung djch th) , th ngui ta thung lc
(nu enzyme cn lc th vic ny cuc ky
quan trong). m cng rt quan trong (chi
c tc dung nui cy b mt), phu thuc
vo thnh phn mi trung b mt.
Khi lua chon mi trung cn ch dn c
thnh phn djnh tnh v djnh luong sao cho
qu trnh sinh tng hop enzyme mong
mun l cao nht. Mun vy ngui ta c
th s dung mt s phuong php:
Phuong php ti uu ha quy hoach thuc
nghim ton phn.
Phuong php ton hoc m hnh ha thuc
nghim.
2.2.4. Cc loai mi trung nui cy vi sinh
vt tng hop enzyme.
C th chia lm 2 loai: mi trung tng hop
v mi trung tu nhin.
Mi trung tng hop: l mi trung bao
gm cc cht vi liu luong xc djnh (qua
tm hiu, nghin cu), chng han ngun
cacbon c th l tinh bt, xenluloza, dung,
axit, ruou, ngun nito v co v hu co.
Loai mi trung ny duoc s dung cho
muc dch nghin cu.[1]
Mi trung tu nhin: thung dng cc
loai ph liu, nguyn liu c cha cc
ngun cacbon, nito, khong, cc yu t
sinh tng hop trung. Mt khc, cc
nguyn liu ny c sn, r tin nn duoc
s dung rt nhiu trong cng nghip sn
xut cc ch phm enzyme t vi sinh vt.
Cc nguyn liu d chun bj lm mi
trung tu nhin bao gm: cm v bt hat
cc, nuc chit ng, djch p hoa qu, rau,
kh du, b ruou, r dung, sn phm
phn hu nm, men bia, tru, li ng. Khi
lua chon s dung mi trung cn ch
dn cc cht c tc dung diu ho sinh
tng hop enzyme, dc bit cc cht cm
ng.[2]
C nhiu yu t nh hung di vi mi
trung nui cy. Nhit d nui cy thng
thung t 25 - 300C. Trj s pH ban du
ca mi trung (ch yu mi trung
nuc) cng c th gy nh hung no d
dn su tao thnh enzyme, nhung khi d
cng cn tnh dn kh nng bin di nhanh
chng chi s d bi vi sinh vt.
2.2.5. Cc phuong php nui cy vi sinh
vt.
Nui cy b mt:
Phuong php ny rt thch hop d nui cy
cc loai nm mc do kh nng pht trin
nhanh, manh, nn t bj tap nhim. Khi nui
nm mc pht trin bao ph b mt hat
cht dinh dung rn, cc khun ty cng
pht trin dm su vo lng mi trung d
duoc tit trng, lm m. i vi mt s
muc dch dc bit, ngui ta nui vi sinh vt
truc tip trn b mt hat gao (sn xut
tuong), hat du tuong (du tuong ln men-
mis) d duoc nu chn trn hat cc cn
sng (lm men thuc bc, men dn tc, lm
tuong).
Ngui ta thung dng cm m, cm gao,
ng mnh. c cht phu gia l tru. Cm,
tru, c b mt tip xc ln, mng, tao
duoc d xp nhiu, khng c nhng cht
gy nh hung xu dn su pht trin ca
nm mc. Ti l cc cht phu gia phi bo
dm sao cho hm luong tinh bt trong khi
nguyn liu khng duoc thp hon 20%, c
th b sung thm ngun nito v co
((NH4)2SO4, (NH4)2CO), photpho, nito
hu co v cc cht kch thch sinh trung
nhu malt, nuc chit ng, nuc loc b
ruou.

Quy trnh cng ngh:
Nui cy, theo di, x l. Chuyn vo
dung cu nui cy Gieo ging vsv Lam
ngui, toi Thanh trng bng nhit Lm
m Trn Nguyn liu
Uu nhuoc dim:
Nng d enzyme tao thnh cao hon nhiu
ln so vi djch nui cy chm sau khi d
tch t bo vi sinh vt. Trong cng nghip
ruou mun dung ho 100kg tinh bt chi
cn 5kg ch phm nm mc b mt nhung
phi cn dn 100lit nm mc chm d loc
b v t bo vi sinh vt.
Ch phm d dng sy kh m khng lm
gim dng k hoat tnh enzyme, ch phm
kh, d bo qun, vn chuyn, nghin nh
hoc s dung truc tip nu khng cn khu
tch v lm sach enzyme.
Tn t nng luong, thit bj, dung cu nui
cy don gin, c th thuc hin qui m gia
dnh, trang trai cng nhu qui m ln dn
20T/ngy.
Nui cy trong diu kin v trng tuyt di
v trong qu trnh nui cy nu c nhim
trng phn no, khu vuc no th chi cn loai
b canh trung phn d.
Tuy nhin phuong php b mt c nng
sut thp, kh co kh ho, tu dng ho, cn
din tch nui ln, cht luong ch phm
cc m khng dng du.
Nui cy chm
Vi sinh vt duoc nui cy trong mi trung
lng vi co cht ch yu trong da s
trung hop l tinh bt. Chi c mt s t
ging vsv dng ngun co cht cacbon l
dung glucoza, saccharoza. Thuc t, trong
mt s trung hop dung ho so b tinh
bt truc khi thanh trng. Khi d dung
maltoza duoc tao thnh l cht cm ng
tt, mi trung thung gim d nht nn
d dng cho qu trnh khuy trn v suc
kh.
Phuong php nui cy b su di hi phi
duoc v trng tuyt di cc khu v sinh
tng hop, thanh trng mi trung dinh
dung, thao tc nui cy, khng kh cung
cp cho qu trnh nui cy.
Cc giai doan ca qu trnh nui cy chm
1 buc gm: chun bj mi trung nui cy,
nui cy nm men ging, nui cy nm
mc sn xut.
Uu nhuoc dim
Phuong php nui cy hin dai d co kh
ho, tu dng ho, nng sut cao, d t
chc sn xut.
C th nui cy d dng cc chng vi sinh
vt dt bin c kh nng sinh tng hop
enzyme cao v lua chon ti uu thnh phn
mi trung, cc diu kin nui cy, enzyme
thu duoc tinh khit hon, dm bao diu kin
v sinh, v trng.
Tuy nhin do thu duoc canh trung v
nng d enzyme thp nn khi tch thu hi
enzyme s c gi thnh cao. Tn din nng
cho khuy trn, nu khng bo dm v
trng s bj nhim hng loat, ton b gy
tn thuong ln.
2.2.6. Tch v lm sach ch phm enzyme.
Mt diu cn ni thm na l enzyme
thung cha cc t bo sinh vt goi l
cc enzyme trong t bo (intracellular),
nhung n cng c th duoc cc sinh vt
tit ra mi trung sng. l cc enzyme
ngoi t bo (extracellular). Enzyme vi sinh
vt thung chit l enzyme ngoai bo.[2]
. Cc phn t enzyme khng c kh nng
di qua mng ca t bo v mng ca cc
cu t ca t bo. Do d d c th chit rt
cc enzyme ni bo, buc du tin l phi
ph v cu trc ca cc t bo c cha
enzyme v chuyn chng vo dung djch.
C th ph v cu trc ca cc t bo bng
cc bin php co hoc nhu nghin vi bt
thy tinh hoc ct thach anh, lm dng ha
bng thit bj dng ha (homogenizator).
Mun tch duoc cc enzyme trong cc cu
t ca t bo, ngui ta cn phi dng cc
yu t vt l v ha hoc khc nhau nhu
sng siu m, dng cc dung mi hu co
nhu butanol, aceton, glycerin, ethyl
acetate... v cht detergent. Cc ha cht
c tc dung tt cho vic ph v cc cu t
ca t bo v trong cc co quan ny thung
cha m.
Sau khi d ph v cc cu trc ca t bo,
enzyme duoc chit bng nuc ct, bng
cc dung djch dm thch hop hoc cc
dung djch mui trung tnh.
C mt s yu t nh hung dn qu trnh
chit rt cn luu . Truc ht d l nhit
d. trnh mt hoat tnh hoc thm ch
v hoat, cn chit rt v tin hnh kt ta
enzyme nhit d thp (t 3 dn 50C).
Cc thao tc phi nhanh. Mt s cht din
ly lm tng qu trnh chit rt enzyme nhu
NaCl, ZnCl2, CaCl2. Tc dung ca chng
cn phu thuc vo phuong php dng khi
chit rt.
loai b mui khong v cc loai
dung... l cc tap cht c phn t luong
thp, ngui ta thung dng phuong php
thm tch (dialysis) di nuc hay di cc
dung djch dm long hoc bng cch loc
qua gel sephadex.

loai b cc protein tap (protein cu trc,
protein tro) v cc cht c phn t luong
cao khc ngui ta hay dng kt hop cc
phuong php khc nhau: phuong php
bin tch chon loc nh tc dung ca nhit
d hoc pH ca mi trung, phuong php
kt ta phn doan bng mui trung tnh
hoc cc dung mi hu co, cc phuong
php sc k (sc k hp phu, sc k trao
di ion), din di, phuong php loc gel.

-Muc dch yu cu: Cc ch phm enzyme
duoc s dung cc dang khc nhau theo
mc d tinh khit (hoat d ring). Trong
mt s trung hop, canh trung nui cy vi
sinh vt c cha enzyme duoc s dung
truc tip dui dang th khng cn tch tap
cht nu chng khng gy nh hung dng
k dn sn phm v quy trnh cng ngh
sau ny (V du: sn xut ruou, nuc chm
thuc vt, da). Cng c khi ngui ta cn s
dung ch phm enzyme tinh khit trong
cng nghip dt, cng nghip mach nha, y
hoc, nghin cu khoa hoc.
Enzyme ni chung rt d bj gim hoat tnh
dui tc dung ca cc tc nhn bn ngoi
do d khi tch v tinh ch enzyme d trnh
su bin hnh protein nh hung ln dn
hoat tnh enzyme cn tin hnh nhanh
chng nhit d thp, d pH thch hop
khng c mt cc cht gy bin hnh
enzyme.
2.2.6.1. Phuong php kt ta
Phuong php kt ta phn doan bng
(NH4)2 SO4 dua trn co s su khc nhau
v kh nng kt ta ca cc protein
enzyme mt nng d mui (tnh theo
phn % nng d bo ha) xc djnh duoc
dng ph bin d loai b buc du protein
tap ca cc djch enzyme. Cc loai mui c
th duoc dng l (NH4)2 SO4, Na2 SO4,
MgSO4 ... ngui ta d nhn thy mui
(NH4)2 SO4 l tt nht v n khng lm hai
m lm n djnh (lm bn) hu ht cc loai
enzyme. Loai mui ny lai r v ph bin.
ha tan ca n lai rt ln (bo ha
767g/l 250C).
2.2.6.2. Phuong php sc k
Cc k thut sc k ct
Djch chit enzyme d duoc loai b phn
ln cc protein tap nhung vn chua dm
bo d dng nht cn thit. Do d djch
chit enzyme duoc tip tuc lm sach bng
phuong php sc k ct.
-Phuong php dng cht ry phn t (loc
gel - gel filtration)
Co s ca phuong php loc gel l dua vo
su khc nhau v kch thuc hnh dang v
phn t luong ca enzyme c trong hn
hop d tch chng ra.
dm bo cho vic tch enzyme duoc
tt, cht ry phn t phi l cht tro, khng
phn ng vi protein enzyme. Cht ny
cng khng ha tan v tuong di bn vi
cc yu t v co hoc cng nhu sinh hoc.
Ngoi ra cht duoc s dung cho muc dch
loc phn t phi l cht khng c tnh dn
hi (khng co) v phi l cht ua nuc
(hydrophyl).


coarse), hat trung bnh (medium), hat mjn
(fine), hat siu mjn, rt mjn (superfine).

-Phuong php sc k trao di ion
Phuong php sc k trao di ion dua vo
su khc nhau v din tch tng s ca cc
protein enzyme. Hay ni cch khc,
phuong php ny duoc dua trn co s ca
phn ng trao di ion gia protein duoc tan
trong nuc hoc dung djch dm long v
cc tc nhn trao di ion. Tc nhn (hay
nguyn liu) trao di ion c th l cht
nhua c tch nhm sinh ion hoc l cht
ionit. y l nhng cht gi tro, khng tan
trong nuc, c bn cht l cellulose hoc
cht gel dextran c lui phn nhnh
(Sephadex, Molselect) hoc l cht nhua
polystirol. Cht gi th ny thung kt hop
vi cc nhm ion ha. Cc cht trao di ion
c cht gi l cellulose, sephadex,
molselect thng thung duoc dng d tch
protein enzyme, cn cc cht trao di ion
c cht gi l polystirol (v du nhu Dowex,
Amberlite) chi dng d tch cc peptid c
trong luong phn t nh hon.
Anionit DEAE - cellulose (diethylamino -
ethyl - cellulose) l dn xut este ca
cellulose.) C2H5

y l cht trao di anion
Nu cht gi l sephadex th chng ta c
cht trao di ion sephadex. l cc loai
DEAE - sephadex v CM - sephadex. Uu
dim ca loai ny l va tch duoc protein
enzyme v kch thuc v v din tch tng
s ca cc protein enzyme. Trung hop
CM - sephadex trn cht gi sephadex c
gn nhm COO-

y l cht trao di cation.
Ngoi ra c cc phuong php :
- Phuong php dng cht hp phu
- Phuong php dng cht hp phu dc hiu
sinh hoc hay l phuong php sc k i luc
(affinity chromatography)

2.2.6.3. Phuong php tch h hai pha
nuc.[4]
Co s k thut ny dua vo su phn b
khc nhau ca hn hop trong dung djch hai
pha khng ha tan vo nhau. Cc h hai
pha dc trung bng cch trn cc h dung
mi vo nhau d tao thnh hai pha ring
bit. H dung mi duoc s dung trong
phuong php ny c th l
polymer/polymer nhu: polyethylene glycol
(PEG) v dextran hoc polymer/mui nhu
PEG v potassium phosphate, sodium
sulphate, sodium phosphate,... Cc protein
v mnh v t bo c kh nng ha tan
khc nhau gia hai pha, v vy phuong
php ny c th duoc dng cho c hai
trung hop: phn tch protein khi mnh
v t bo v phn chia cc enzyme trong
sut qu trnh tinh sach protein. H hai pha
nuc duoc xem l phuong php tt cho
vic phn tch v tinh sach cc hn hop, v
phn tch cht lng c mt d khc nhau
d dng hon phn tch cc cht rn ra khi
cc cht lng. So vi cc phuong php tinh
sach khc th h hai pha nuc c mt s
uu dim hon nhu: c d ha tan trong
nuc ca hai pha ln (70-80%), dat d tinh
sach cao, hiu sut cao, d dng s dung
quy m ln v dc bit polymer duoc ti
s dung.
Su phn tch dat hiu qu cao ty thuc
vo cc thng s nhu khi luong phn t
v din tch ca di tuong nghin cu,
nng d v khi luong phn t ca cc
polymer, nhit d, pH, thi gian, luc ion ca
hn hop v su hin din ca cc loai mui
da ha trj nhu phosphaste hoc sulphate...

Hnh 2.9. Phuong php tch h hai pha
nuc
2.3. Enzyme c djnh.
Enzyme c djnh (hay enzyme khng tan) l
enzyme c su tham gia hoat dng trong
mt khng gian bj gii han. Su gii han
hoat dng vn linh hoat ca enzyme bng
cch gn n vo mt pha cch ly tch ri
khi pha lng tu do v d n vn c kh
nng tip xc duoc vi cc phn t co
cht, effector hay inhibitor (cht c ch).
Pha gn enzyme thung khng tan trong
nuc nhung cng c th l cc polyme uu
nuc
Loi ch ca vic s dung enzyme c djnh.
-Gim gi thnh do enzyme duoc s dung
lp lai nhiu ln vi cng mt kiu phn
ng xc tc, ch phm bn hon trong cc
diu kin pH, nhit d, p sut thm thu
ti uu, tc d phn ng ln, d t chc sn
xut mc d tu dng ha cao.
-Ch tao enzyme tuong di d, du tu xy
dung v sn xut tuong di t, sn phm
phn ng khng ha ln ln vi enzyme,
c th d dng t chc sn xut cc sn
phm ln men bng enzyme ngoai bo
nhu: ruoc etylic, axit hu co, axitamin,
vitamin.
Mt s phuong php ch yu tao enzyme
c djnh
-Microencapsulation (gi enzyme vo trong
cuc nh)
uoc nghin cu bi Chang et al(1967-
1968)
Ci mng polymer thm thu dy 200A0
(xenluloza, polysacarit, phi tinh bt) tao
thnh hat dng vng 10- 12M cha c
phn t enzyme bn trong. Lp mng ny
cho php co cht v sn phm phn ng
eyme duoc qua lai tu do nhung cc phn
t enzyme khng th qua lai duoc v cc
phn t qu ln nhu : polysaccarit, protein
cng khng th qua lai mng duoc do d
khng th thuc hin phn ng enzyme
duoc.
Ngoi ra cn c cc phuong php:
-Lin kt enzyme vo cht mang khng tan
(silman v katchalski-1966)
-jnh vj enzyme trong pha lng ca h
thng 2 pha (Reese and Mandels 1958)
-Gi enzyme bng mng siu loc.

Cc thit bj sn xut enzyme c djnh

Figure 2.11. Sketch of the pilot-scale kinetic
mixer

Figure 2.2 Photograph of kinetic mixer after
installation at BNL
moisture contents but could not be
remedied with the current mixer design.

collection vessel, and rectangular HEPA
filter box are clearly discernable in the
photograph.

Figure 2.12. Schematic of kinetic mixer off
gas system.

Figure 2.13. Photograph of kinetic mixer off
gas system.

Figure 2.14. Photograph of kinetic mixer
with end flange removed for access to
mixing blades.

Figure 2.15. Photograph of mixing blade
sleeve.

Figure 2.16. Photograph of Plexiglass
containment enclosure.

Figure 2.17. Photograph of enclosure
during smoke bomb leak test.

CHUONG . KT QU V THO LUAN
3.1. Kh nng sinh tng hop enzyme
Protease t vi sinh vt
Kim tra su tng trung vi sinh vt: ly v
du minh hoa kim tra bng ha cht.
Bng 3.1. Kh nng sinh tng hop protease
ca XKDNW10 theo thi gian 500C.[5]

Thi gian nui cy Vng phn gii

36 gi

48 gi

60 gi

72 gi

84 gi


T dung knh vng phn gii d thj v kh
nng sinh enzyme protease ca chng
XKDNW10 d duoc xy dung (hnh 3.1.).

t(h) 24 36 48 60 72 84
D(mm) 0 9 15 22 29 29
Ch thch: t(h)- Thi gian nui cy, d(mm)-
ung knh vng thy phn
3.2. ng dung enzyme Protease vo trong
cng nghip.
Protease duoc s dung rng ri trong
nhiu ngnh cng nghip nhu cng nghip
thuc phm, cng nghip nhe, cng nghip
duoc phm v nng nghip.
Trong cng nghip ch bin thjt, protease
duoc dng lm mm thjt nh su thy phn
mt phn protein trong thjt, kt qu lm cho
thjt c mt d mm thch hop v c vj tt
hon. Protease duoc s dung d lm mm
thjt v tng huong vj thjt. (ngm thjt vo
dinh dung protease pH v nhit d xc
djnh- phuong php ny ph bin v thun
loi nht; Tm hn hop lm mm thjt
(enzyme, mui, bt ngot). Tim dung djch
enzyme vo thjt; tim dung djch enzyme
vo con vt truc khi git m). S dung
protease d sn xut djch dam: t
Streptomyces fradiae tch duoc ch phm
keratineza thu phn duoc keratin rt c
gi trj d sn xut djch dam t da, lng v.
Nu dng axit d thu phn s mt di hon
ton cc axit amin cha luu huynh, nu
dng kim d thu phn s bj raxemic ho
(chuyn dang L sang D lm gim gi trj
sinh hoc ca axit amin). thu phn su
sc v trit d protein (trong nghin cu,
ch tao djch truyn dam y t) cn dng cc
protease c tnh dc hiu cao v tc dung
rng, mun vy ngui ta thung dng phi
hop c 3 loai protease ca 3 loi: vi khun,
nm mc, thuc vt vi t l tng cng 1-
2% khi luong protein cn thu phn. Uu
dim ca vic thu phn protease bi
enzyme l bo ton duoc vitamin ca
nguyn liu, khng tao ra cc sn phm
phu, khng lm sm mu djch thu phn.
Trong ch bin thu sn: Khi sn xut nuc
mm (v mt s loi mm) thung thi
gian ch bin thung l di nht, hiu sut
thu phn (d dam) lai phu thuc rt nhiu
dja phuong, phuong php gi nn, nguyn
liu c. Nn hin nay quy trnh sn xut
nuc mm ngn ngy d duoc hon thin
trong d s dung ch phm enzyme thuc
vt (bromelain v papain) v vi sinh vt d
rt ngn thi gian lm v ci thin huong vj
ca nuc mm. Tuy nhin vn cn mt s
tn tai cn phi hon thin thm v cng
ngh.
Trong cng nghip sa, protease duoc
dng trong sn xut phomat nh hoat tnh
lm dng tu sa ca chng. Protease t
mt s vi sinh vt nhu A. candidus, P.
roquerti, B. mesentericus,. duoc dng
trong sn xut pho mt [1]. Trong cng
nghip sn xut bnh m, bnh quy...
protease lm gim thi gian trn, tng d
do v lm nhuyn bt, tao d xp v n
tt hon.
Trong sn xut bia, ch phm protease c
ngha quan trong trong vic lm tng d
bn ca bia v rt ngn thi gian loc.
Protease ca A. oryzae duoc dng d thy
phn protein trong hat ng cc, tao diu
kin x l bia tt hon [6].
Trong cng nghip da, protease duoc s
dung lm mm da nh su thy phn mt
phn protein ca da, ch yu l collagen,
thnh phn chnh lm cho da bj cng. Kt
qu d loai b khi da cc cht nht v lm
cho da c d mm do nht djnh, tnh cht
d duoc hon thin hon sau khi thuc da.
Truc dy, d lm mm da ngui ta dng
protease duoc phn lp t co quan tiu
ha ca dng vt. Hin nay, vic dua cc
protease tch t vi khun (B. mesentericus,
B. subtilis), nm mc (A. oryzae, A. flavus)
v xa khun (S. fradiae, S. griseus, S.
rimosus...) vo cng nghip thuc da d
dem lai nhiu kt qu v dn dn chim
mt vj tr quan trong [2] .
Trong cng nghip dt: Proteinase vi sinh
vt duoc s dung d lm sach to tm, ty
to nhn tao (cc soi nhn tao duoc bng
cc dung djch cazein, gelatin) d soi duoc
duoc bng, d nhum. Protease c tc
dung thy phn lp protein serisin d lm
dnh bt cc soi to tu nhin, lm bong v
tch ri cc loai to tm, do d lm gim
luong ho cht d ty trng.
Ngoi ra, protease cn duoc ng dung
rng ri trong rt nhiu ngnh khc nhu:
- iu ch djch dam thy phn dng lm
cht dinh dung, cht tng vj trong thuc
phm v sn xut mt s thc n king.
- Protease ca nm mc v vi khun phi
hop vi amylase tao thnh hn hop
enzyme dng lm thc n gia sc c d
tiu ha cao, c ngha ln trong chn
nui gia sc v gia cm.
- iu ch mi trung dinh dung ca vi
sinh vt d sn xut vaccine, khng sinh,.
- Sn xut keo dng vt, cht git tng hop
d git ty cc cht bn protein, sn xut
m phm,.
KT LUAN
Trong qu trnh thuc hin bi semina:
"Cng ngh sn xut enzyme Protease t
vi sinh vt ng dung trong cng nghip
chng ti d dat duoc cc kt qu sau:
-Hiu ngha,vai tr ca VSV trong sn
xut v di sng x hi, dc bit trong
tuong lai khi cng ngh sinh hoc pht trin.
-Nm duoc dc tnh sinh l ca vi sinh vt.
-Bit duoc mt s nhm vi sinh vt chnh
c th sn xut enzyme Protease ng dung
trong cng nghip.
T LEU THAM KHO
TNG VET
1. Trn Xun Ngach (2007), Cng ngh
enzym, Trung ai Hoc Bch Khoa
Nng.
2. Nguyn Trong Cn, Nguyn Thj Hin,
Thj Giang, Trn Thj Luyn (1998), Cng
ngh enzyme, Nh xut bn nng nghip
thnh ph H Ch Minh.
3. Luong c Phm (2004), Cng ngh vi
sinh vt, Nh xut bn Nng nghip, H
Ni.
4. Luu Thj Nguyt Minh(2007), "Phn tch
Protease ca Bacillus Subtilis bng h hai
pha Polyethylene glycol/ Potassium
phosphate, n tt nghip.
5. Pham Trn Vnh Ph (2007), " Nghin
cu mt s chng vi sinh vt ua nhit t
qu trnh phn hy tu nhin rc thi sinh
hoat ca thnh ph Nng, n tt
nghip.
6. L Xun Phuong (2001), Vi sinh vt
cng nghip, NXB Xy dung, H Ni.

T LEU MANG

1. http://www.zuova.cz/informace/nrlpab23.
php
2.http://en.wikipedia.org/wiki/Protease 3.htt
p://vietsciences2.free.fr/khaocuu...n_baotu0
03.jpg
4.http://images.google.com.vn/imgres?i...sc
hmittelvergl
5. http://faculty.ircc.edu/faculty/tfis...sformati
on.jpg