Professional Documents
Culture Documents
GIO TRNH
CNG NGH SINH HC THC PHM II
THI LNG 45 TIT
NNG 2007
M U
Cng ngh sinh hc l mt ngnh khoa hc ang c th gii quan tm. Gi
tr ca cng ngh sinh hc l ch l mt cng c c th p dng cho nhiu ngnh
kinh t nh sn xut lng thc thc phm, chn nui th y, cng nghip dc v
cng nghip ho hc, chuyn ho sinh khi thnh nng lng, x l ph liu v ph
liu cng nng nghip, phng chng nhim v v sinh mi trng
- Cng ngh sinh hc gip cho cc chng trnh ci thin nng nghip,
ngh vn v ngh rng tng nng sut cy trng, tng cht lng dinh dng ca
nng phm, chn ging chng chu vi su bnh v vi thi tit t ai khng thun
li v to ra cc ging thch nghi vi cc iu kin th nhng v kh hu nht nh.
ng thi n cng to iu kin duy tr mt s a dng di truyn rng gia cc
ging cy trng v gi gn cc ngun gen c to nn t cc h hng hoang di
ca chng. Tnh a dng di truyn c th hin thng qua mt s lng cc ln cc
kiu kt hp gen c trong mt s c th ca mt loi v thng qua s khc nhau v
cc tnh trng ca cc ging trong cng mt loi: kiu sinh trng, tnh khng su
bnh, tnh khng vi ngoi cnh (sng mui, hn, nng) v nng sut. Sau khi
nghin cu kt qu ca nhiu t hp lai mt cch cn thn v nghim tc, cc nh
chn ging da v tnh a dng di truyn chn ra cc dng c tnh trng mong
mun. thnh cng, nh chn ging phi c c trong tay vn di truyn cng ln
cng tt. Vn di truyn ny bao gm cc ging cy trng, cc ging chng chu, cc
ging a phng (cc ging ny thng b b qun v do nng sut thp nhng n rt
qu v c tnh chng su bnh v cc iu kin ngoi cnh khng thun li). Trong
vn di truyn cn phi k n cc cy hoang di c tc dng tng sc sng cho cc
cy ging trng.
Tnh a ng di truyn l nhn t bo m cy trng khng b cc tai bin kh
hu hoc su bnh tiu dit hon ton. Cy trng cng thun nht th cng d b hi
khi c tai ho.
- Cng ngh sinh hc cng c ng dng rng ri trong lnh vc thc phm
v dinh dng. Ngnh cng ngh ln men l mt b phn ca cng ngh sinh hc
sn xut ra nhiu sn phm rt th v cho ngnh thc phm. Hoc vic sn xut nm
men cng c mt ngha rt quan trng. Ngi ta c th s dng mt lng nh nm
men b sung protein, vitamin v cc cht khong cho thc phm. Ngoi ra, sinh
khi nm men l ngun thc n b sung trong chn nui rt c hiu qu.
- Cng ngh sinh hc gip cho chn nui v th y to ra c nhng ging
nui mong mun v sn xut ra cc loi vacxin phng chng bnh tt cho cc vt
nui. V d: vic cy chuyn hp t b to ra c ging b tt, c sc chu ng
cao. K thut cy chuyn c thc hin nh sau: gy s rng trng mt con b ci
c cc c im m ngnh chn nui cn n v em th tinh nhn to bng tinh
trng ca mt con b c c nhng c im nh ngi ta mong mun. Cc hp t
hay phi c thu nhn bng cch ra d con. Lm ng lnh phi trong nit lng
-179oC v c th vn chuyn trng thi ny: 1000 phi ng lnh khng nng qu
50kg. Phi c cy vo b ci cha h. B con pht trin ln t cc phi ny s
ra i trong mi trng sng sau ny ca chng v khng phi ng u vi nhng
s bt li ca mi trng m cc sc vt nhp hay gp phi. S bo v bng cc
khng th ca b m v nh b sa ca b m m sau khi ra i b s sinh c th
chu ng tt hn i vi cc loi bnh tt thng thng.
- Cng ngh sinh hc cn tch cc gip ngnh y t bo v sc kho ca
cng ng. Ngi ta ni rng y hc d phng (v lu di hn l y hc d bo da trn
hiu bit v c im di truyn mi c th) s mang li li ch nhiu hn cc phng
php iu tr. Mt lnh vc ca cng ngh sinh hc c th gp phn quan trng trong
gii quyt cc vn y t, trong khun kh mt chnh sch u tin cho y hc d
phng, l ci tin cc vacxin hin c v ch to ra cc vacxin mi. ng thi n
cn gip ch trong vic sn xut cc loi khng sinh, vitamin v cc thuc cha bnh
khc. Nhng nm gn y, nh vo k thut di truyn ngi ta tm cch tch chit
cc hot cht ca thc vt bc cao lm vt liu xut pht cho hng lot loi thuc.
- Cng ngh sinh hc cn gp phn trong vic sn xut ra nng lng nh: sn
xut cn bng con ng ln men, chng trnh biogasv chnh cc ngun nng
lng ny li i phc v cho cc qu trnh sn xut khc.
- Cng ngh sinh hc cn tham gia vo vic chuyn ho cc cht v ngn chn
s nhim mi trng. Sn phm ph v cc cht thi cha hydratcacbon c th
c chuyn ho bng cch ln men nh cc vi sinh vt thng thng hay bng cc
qui trnh cng ngh sinh hc. Hoc cc k thut ti t hp AND cng s gp phn
tch cc tch c cc ging vi khun thch hp nht cho vic ti u ho nhng s
chuyn ho . V d: chuyn gen m ho cc enzym xenluloza v hemixenluloza ca
Clostridium thermocellum thnh nhng loi Clostridium khc c th iu khin c
s chuyn ho xenluloza v hemixenluloza thnh etanol, axeton, butanol, axt axetic
v axt lactic. Dng mt vi ging a nhit Clostridium (toop= 65-75oC) c li l ct
gim c chi ph trong vic chng ct v s h c gi thnh sn phm.
ngn chn s nhim mi trng ngi ta c th dng cc loi vi sinh vt
khc nhau. Cc k thut ti t hp AND to ra nhng chng vi khun c th phn
hu v hp th mt s ln cc cht do cng nghip ho cht thi ra.
Nh vy cng ngh sinh hc c lin quan n nhiu lnh vc v bao gm cc
ngnh nh:
- Cng ngh di truyn
- Cng ngh nui cy m v t bo
- Cng ngh enzyme
- Cng ngh vi sinh vt
Trong hc phn ny s nghin cu v mt s phn ca cng ngh vi sinh vt.
Cc qu trnh vi sinh c s dng rng ri trong cc lnh vc khc nhau ca nn
kinh t. Nhng thnh tu khoa hc k thut v sinh ho cho php to ra nhng qu
trnh sn xut c nng sut cao da trn cc phng php cng ngh c iu
chnh c mt s sn phm thc phm, chn nui, thuc cha bnh v cc cht hu
c.
2. Cc cht trao i bc 1
V d: axt amin, nucleotit, vitamin, axt xitric.
Cc cht trao i bc mt l nhng vin gch cu trc c trng lng
phn t thp ca cc cao phn t sinh hc ca t bo cht: axt amin, nucleotit,
nucleozit, ng, axt bo, vitamin Ngoi ra cc sn phm trung gian ca qu
trnh trao i cht (cc axt hu c trong chu trnh ATC) cng l cc cht trao
i bc 1. Cc c ch iu ho pht trin trong qu trnh tin ho bo m sao
cho cc cht trao i bc 1 ch c tng hp n mc cn thit.
3. Cc cht trao i bc 2
V d: khng sinh, alcaloit
Cc cht trao i bc 2 l nhng cht trao i c trng lng phn t
thp, khng gp mi c th, s phn b ca chng ch gii hn nhng n
v phn loi nht nh. Chng khng c chc nng chung trong trao i cht
ca t bo v t bo cng c th tn ti m khng cn n chng. Tuy nhin,
cc cht trao i bc 2 c th c ngha vi s sinh trng v cc c th sn
sinh ra chng. Chng hn mt s cht trao i bc 2 c vai tr trong s hp th
st khi thiu nguyn t ny. Thng cc cht bc 2 c to thnh khi s sinh
trng kt thc. Cc chng tn ti trong t nhin thng ch to thnh rt t
cht trao i bc 2, nhng cht ny c tch lu trong t bo hoc thi ra
ngoi.
4.Cc enzym
V d: enzym ngoi bo: proteaza, amylaza
enzym ni bo : asparaginaza, penixilinaza.
T bo vi sinh vt cha khong 1000 enzym khc nhau. Mt s enzyme
ch c mt vi s lng vi phn t nhng nhiu enzym c mt vi s lng
ln. Ch nhng enzym chu trch nhim phn hu cc c cht khng ho tan
nh tinh bt, xenlulo, proteinmi c tit t t bo vo mi trng. Vi sinh
vt c kh nng s dng cc c cht khc nhau cho sinh trng v thch ng
vi cc iu kin sinh trng rt khc nhau. Trong s gii hn ca th tch t
bo ch tng hp nhng enzym m n cn. Trong cng ngh sn xut enzym
cn phi iu khin trao i cht sao cho enzym m ta mong mun c tng
hp cng nhiu cng tt.
1.1.3 Cc sn phm ca s chuyn ho cht
V d: s oxy ho khng hon ton to thnh axit axetic soboza.
Trong qu trnh chuyn ho cc t bo hot ng nh nhng h thng
xc tc cho mt hoc nhiu bc chuyn ho cht. V mt l thuyt nhng phn
ng ny cng xy ra nh cc enzym c lp, tuy nhin con ng ny khng th
thc hin c hoc khng kinh t (v d vi cc phn ng enzym ph thuc
nng lng). Cc enzym chuyn ho cc cht theo cch rt c hiu. V d: s
ng thi
Thi gian
Khng ng thi
Thi gian
Cc enzym cu trc
4
Glucoza
Kim ch d ho
Kim ch bng sn phm
cui cng
E.1
E.2
E.3
E.4
c ch bng sn phm
cui cng
S 1: Cc nguyn tc ca s iu ho enzyme
* Trong c ch 1 sn phm cui cng ca 1 qu trnh sinh tng hp gy
ra s c ch qu trnh tng hp ca chnh n. y, sn phm cui cng d
c hnh thnh trong t bo hay c thu nhn t mi trng dinh dng,
iu cng ngha. Trong c ch ny, sn phm cui cng ni chung nh
hng n enzyme u tin ca chui sinh tng hp. Enzym c tnh quyt nh
ny l 1 protein d lp th. N c c im l thay i cu hnh khng gian khi
Hot ng
Cht nh hng
d lp th
Cht nh hng
d lp th
Th t bin
Khng hot ng
Dng hoang di
Gen
iu
khin
khng phong to s
truyn thng tin tng
hp enzym
Cc
gen
cu
trc
cht kim ch
hot ng
Cht kim ch
hot ng
Th t bin
Tinh bt glucose
amylaza
Qt. ngoi bo
Cc enzym
i vi cht bo:
Cht botriglyceritaxt bopalmityl-CoA axetyl-CoA
2. Nguyn liu cung cp ngun nit
Vi sinh vt cn nit xy dng t bo v tt c cc thnh phn quan
trng ca t bo u c cha nit (protein, axt nucleic, enzym).
Axt amin to ra protein. Cc axit amin trong t bo vi sinh vt c to
thnh do qu trnh trao i cacbon v nit.
Nit trong khng kh rt phong ph nhng n bn vng v mt ho hc
v ch c cc vi sinh vt c nh nit mi c kh nng ng ho chng. Cn cc
ngun nit dng trong ln men ca cng ngh vi sinh vt l cc hp cht nit
v c v hu c.
Cc axit amin thng khng c vi sinh vt s dng trc tip ngay m
phi qua 2 loi phn ng: hoc kh amin hoc kh cacboxil.
Kh amin:
.deza min aza
R-CH-COOH + O2 E
R-C-COOH + NH3
NH2
E.
R-CH-COOH
R-CH2-NH2 + CO2
NH2
G mi 500 gi
Heo 1000 gi
Tru b 2000 gi
Nh vy trong cng mt thi gian nu nui cy vi sinh vt thu nhn
sinh khi s thu c mt khi lng cao hn rt nhiu so vi cc sinh vt
khc.
3. Hm lng protein cao:
Hm lng protein ph thuc vo chng v chu nh hng nhiu iu
kin nui cy. Ni chung hm lng protein vi sinh vt t 50-60%.
4. Cht lng protein cao:
nh gi cht lng protein ngi ta hay quan tm n hm lng axit
amin khng thay th. Tiu chun ny cng mang tnh cht loi. Thnh phn axit
amin ca protein vi sinh vt ging nh trong tht v sa. Protein vi sinh vt giu
lizin nhng cc axit amin cha lu hunh li thp.
5. Kh nng tiu ho ca protein:
Kh nng tiu ho protein vi sinh vt b hn ch bi cc thnh phn nit
phi protein (nh axt nucleic, peptit ca thnh t bo). Tuy nhin nu protein
c tch khi t bo th khi quan tm. Do bn cht thnh t bo l tiu
chun la chn vi sinh vt dng trong sn xut protein vi sinh vt.
6. S an ton v c t:
Cc vi sinh vt gy bnh hoc c cha cc thnh phn gy c nghi ng
th khng c dng sn xut protein. Yu cu hm lng axt nucleic phi
thp, hm lng ca n cng cao th cng lm gim gi tr ca protein. Khi tiu
ho cc axt nucleic s phn hu tip thnh cc nucleotit, sau li c phn
hu tip thnh ademin hoc guanin v cui cng thnh axt uric. Trong c th
ngi khng c enzym urinaza do uric khng chuyn ho tip. S tch t
axt uric s gy nn bnh thp khp, to ra si thn, si bng quang do ho
tan thp ca axt ny. Lng axt nucleic hp th qua dinh dng khng c
qua 2g/ngy.
ng vt vn ny khng quan trng v chng c kh nng ng ho
axt uric. Ta c th lm gim lng nucleic bng nhng bin php sau:
- Gim mnh tc sinh trng.
- Chit rt ARN=dung dch NaCl 10% nng.
- Thu phn ARN bng kim v tch protein ho tan trong bng kt
ta.
- Phn hu ARN bi enzym nucleaza a vo hoc ca bn thn t bo.
Dng phng php gy chong nhit theo 3 bc: u tin cc t bo c un
nng ln 68oC trong 5 giy. Sau 52,5oC trong 2gi v cui cng 5556oC trong 1h. Vic x l ny lm bin tnh riboxom v hot ho cc
ribonucleaza. Cc sn phm thu phn c tch khi t bo, hm lng
6. X l nm men:
Tu theo yu cu ca sn phm (t hoc kh) m qu trnh x l khc
nhau. thu nhn nm men t th tin hnh ra sinh khi thu c ri em
p. Nm men t kh bo qun v thi gian bo qun khng lu.
bo qun lu th cn phi sy kh nm men. Khi sy nm men ch
khng nhit sy vt qu 300C.
3.2.3 Sn xut men nc:
1. Nguyn liu: bt, malt hoc ch phm amylaza ca nm mc.
2. Mi trng nhn ging: bttrn nc (t l 1:3)nu chinlm
ngui 48-50oCb sung ch phm amylaza (3% malt hoc 0,8 1% ch phm
enzym nm mc nh Asp.awamori hoc Asp.oryzae v dch ging vi khun.
Gi 8-14 gi, cho n khi mi trng t ti 11 12o axt (1o=1ml NaOH
1N/100ml) th a vo cy nm men.
3. Dch ging vi khun: ly nc malt c nng 12oBx thm 1 t
CaCO3. Cy vi khun Lactobacterium Delbruckii v gi nhit 50 52oC
trong 12 ngy. Trc khi a vo s dng phi nhn ging tip.
4. Cy nm men: mi trng sau khi axt ho c lm ngui n 28o
30 C ri tip ging nm men nui cy trc vo (t l 510%), gi nhit
ny 1415 gi khng sc kh hoc sc kh gin on. nhit ny vi
khun lactic ngng tch t axt. Nm men s dng ng v axt tng trng
sinh khi.
Trong mi trng c axt nm men s pht trin tt v hn ch cc vi sinh
vt l.
6. X l nm men:
Tu theo yu cu ca sn phm (t hoc kh) m qu trnh x l khc
nhau. thu nhn nm men t th tin hnh ra sinh khi thu c ri em
p. Nm men t kh bo qun v thi gian bo qun khng lu.
bo qun lu th cn phi sy kh nm men. Khi sy nm men ch
khng nhit sy vt qu 300C.
3.2.3 Sn xut men nc:
1. Nguyn liu: bt, malt hoc ch phm amylaza ca nm mc.
2. Mi trng nhn ging: bttrn nc (t l 1:3)nu chinlm
ngui 48-50oCb sung ch phm amylaza (3% malt hoc 0,8 1% ch phm
enzym nm mc nh Asp.awamori hoc Asp.oryzae v dch ging vi khun.
Gi 8-14 gi, cho n khi mi trng t ti 11 12o axt (1o=1ml NaOH
1N/100ml) th a vo cy nm men.
3. Dch ging vi khun: ly nc malt c nng 12oBx thm 1 t
CaCO3. Cy vi khun Lactobacterium Delbruckii v gi nhit 50 52oC
trong 12 ngy. Trc khi a vo s dng phi nhn ging tip.
4. Cy nm men: mi trng sau khi axt ho c lm ngui n 28o
30 C ri tip ging nm men nui cy trc vo (t l 510%), gi nhit
ny 1415 gi khng sc kh hoc sc kh gin on. nhit ny vi
khun lactic ngng tch t axt. Nm men s dng ng v axt tng trng
sinh khi.
Trong mi trng c axt nm men s pht trin tt v hn ch cc vi sinh
vt l.
Glucoza
Ribozo-5- P
Glucozo-6- P
Sixtein
Eritrozo-4- P
3-P-glyxerat
Serin
Pyruvat
CH3COCOOH
Glyxin
-xetoizovalerat
Alanin
Valin
Lxin
+CO2
Oxalatxetat
(HOOCCOCH2COOH)
Asparagin
Aspartat
Fumarat
Xitrat
-xetoglutarat
Lyzin Treonin Methionin
Izolxin
Glutamat
Glutanin Prolin Arginin
COOH
3. C ch:
Glucoza
Pyruvat
Oxalaxetat
1
Aspactat
-Aspactyl-photphat
Aspactat--semialdehyt
3
Lyzin
Homoxerin
Treonin
Methionin
Izoleuxin
S 3: C ch tng hp lyzin ca Corynebacterium glutamicum
1 -Enzym aspactokinaza; 2 -Enzym homoxerindehydrogenaza;3 -Enzym
dihydropicolinat-syntetaza.
Lyzin l mt axit amin thuc h aspactat v c tng hp qua mt con
ng trao i cht phn nhnh m qua homoxerin, metionin, treonin,
izolxin cng c to thnh.
Nhng ng chm chm biu din s c ch bi sn phm cui cng.
chng hoang di lyzin v treonin cng gy ra mt s c ch phi hp (E) i
vi aspactokinaza(1). Do khuyt homoxerindehydrogenaza(2) m khng c s
to thnh treonin. Dihydropicolinat-Syntetaza(3) khng mn cm d lp th nn
s c ch bi sn phm cui cng b trit tiu v c s tng hp tha lyzin
(50g/l).
4. Ln men :
Ngun hydratcacbon thch hp cho tng hp lyzin l glucose, fructose,
maltose v saccarosse. Cc ng nh lactose, rafinose, pentose cc chng sinh
lyzin khng ng ho c. Cho nn ngi ta thng s dng cc loi nguyn
liu nh r ng, dch thu phn t tinh bt (ng, sn) lm mi trng sn
xut lyzin.
Nng ng trong mi trng ln men khong 10-12%. Trong qu
trnh ln men, nhm tng hiu sut thu hi lyzin c th b sung thm ng
nng cao nng ng ln ti 25% (nhng cng thn trng v c khi hiu
3. C ch:
Khi s dng axit antranilic lm tin cht th c ch ca qu trnh nh sau:
COOH
NH2
NH
indol
axit antranilic
COOH
CH2CHCOOH+H2O
NH2
piridoxalfotfat
HOCH2CHNH2+
serin
NH
NH
Triptophan
S 4: C ch chuyn axit antranilic thnh triptophan
4. K thut ln men:
Qu trnh sn xut chia lm hai giai on: thu nhn sinh khi v chuyn
ho tin cht nh s gip ca sinh khi thu nhn c.
Giai on u khng khc my so vi qu trnh nui cy cc loi vi sinh
vt ging khc. Tc l qu trnh cng i t ng ging gcng nghimbnh
tam giclng sinh khi thu c ph thuc vo phng php nui cy
ging. H thy rng nu lng ging a vo ln men cng ln th mc
chuyn ho ca axt antranilic cng ln.
Ngun cung cp cacbon cho ging pht trin l sacaroza hoc mt r (vi
hm lng t 6,3-20%), ngun cung cp nit l ur vi lng t 0,5-1%. Ngoi
ra cn b sung thm (%): K2HPO4-0,01, MgSO4-0,005,CaCl2-0,01. Bo m
mi trng c pH=7,58 v v trng. Tin hnh nui ging nhit
28300C, thi gian nui ca mi cp khng qu 24 gi. Yu cu ging t 35
gam sinh khi kh/1lt mi trng.
Giai on hai c thc hin trong thit b ln men. u tin chun b
mi trng nui ging trong thit b ln men v chuyn men ging nui
giai on 1 vo. Tip tc nui ging trong 24 gi nhit 28300C v thng
kh khng t hn 7g O2/l.h. Nu trong qu trnh nui ging c xut hin bt th
phi dng du ph bt.
Sau 24 gi nui ging cho vo thit b ln men dung dch axt antranilic
5% trong ru v dung dch amoniac 50% ln men. Lc ny thng kh bo
m cung cp 34 gam O2/l.h.
Sau khi cho dung dch axt vo c 3-4 gi th b sung thm dung dch
r ng 25%. Tip theo, c sau 12 gi th b sung dung dch r ng, sau 6
gi th b sung dung dch ammoniac v axit antranilic mt ln. Tng thi gian
ca giai on 2 l 120140 gi.
Qu trnh ln men (giai on 2) thc hin pH ca mi trng 8,0 v
nhit ln men khong 300C.
3. Thu nhn sn phm:
Dung dch sau ln men c cha 7,812,5% cht kh, trong c
0,30,5% triptophan. Phn ln triptophan (8588%) nm trong pha lng. Do
thu nhn ch phm tinh khit th phi tinh ch dch lc. Cn nu dng cho
chn nui th s dng dng c c c c sinh khi nm men.
lm thc n gia sc th dch sau ln men em c c cn li 1/3 th
tch, sau em sy trong thit b sy phun (to=110120oC) thu nhn bt
dng b sung cho thc n gia sc. Trong thnh phn ca bt ny gm c:
- Cht kh 90%
- Protein
4854%
- Triptophan 13%
- Vitamin (mg/kg):
+ B1 : 1518,5
+ B2 : 24,532,5
+ PP : 620680
- Axit amin khc 6%. Trong c cha nhiu cc axit amin nh: lyzin,
histidin, treonin, xerin, glutamate, prolin, glyxin, valin, tirozin, phenilalanin,
acparagin, acginin
thu nhn ch phm triptophan tinh khit th phi qua cc bc sau:
- Tch sinh khi bng li tm
- Dng HCl axit ho n pH = 1 v tch cc kt ta
- Tin hnh trao i ion
- Nh hp ph nh dung dch ammoniac 5%
- C c dung dch nhit 60700C
- Lm lnh n 460C tr tinh
- Tch tinh th triptophan
- Ra cc tinh th bng ru etylic
- Sy cc tinh th nhit 600C
Cc ph liu ca qu trnh tinh ch ny c th dng cho chn nui.
4.3.3.2 Thu nhn nh cc chng t bin:
S thu nhn triptophan cng nh tirozin v phenilalanin c th thc hin
trn cc chng t bin ca E. coli hoc Bacillus subtilis. Qu trnh tng hp
tirozin, triptophan v phenilalanin i theo s sau:
Glucose
Axit fotfoenol pyruvic
Eritrozo-4- P
Axit skimic
Axit corismic
Axit antranilic
Triptophan
Axit prephenic
Axit phenylpyruvic
Axit hydroxyphenylpyruvic
Phenylalanine
Tirozin
S 5: C ch tng hp triptophan, phenilalanin, tirozin trong
t bo vi sinh vt
tng hp tha triptophan ngi ta dng chng e. coli tr dng tirozin
v phenylalanine.
Glucose
Axit fotfoenol pyruvic
Eritrozo-4- P
Axit skimic
Axit corismic
Axit antranilic
Triptophan
Axit prephenic
Axit phenylpyruvic
Axit hydroxyphenylpyruvic
Phenylalanine
Tirozin
S 5: C ch tng hp triptophan, phenilalanin, tirozin trong
t bo vi sinh vt
tng hp tha triptophan ngi ta dng chng e. coli tr dng tirozin
v phenylalanine.
5.2.1 ng vt:
Enzyme nm trong cc c quan ni tng ca ng vt:
- Tu tng: cha nhiu enzyme amylaza, maltaza, proteaza, esteraza,
lipaza.v.v.
- D dy cha nhiu enzyme pepxin, cc enzyme tiu ho (trong ngn th
t ca d dy b c 1 loi vi khun c kh nng sinh tng hp xenlulaza), trong
d dy ca b v ngh c cha enzyme rennin lm ng t sa.
- Rut: cha ch yu l enzyme enterokinaza c kh nng ph v cc lin
kt peptit.
- Gan: l c quan cha rt nhiu enzyme.
Khng th thu enzyme t ng vt trn qui m cng nghip c v thi
gian nui ng vt qu di nn khng kinh t.
5.2.2 Thc vt:
Ngi ta c th thu c mt s enzyme trong thc vt thng ng nh
thu papain t u , bromelain t da, amylaza t malt, plazmin (c papain) t
h ficus (sung, v).
Trong cc h u c enzyme ureaza. Trong thuc l c cha enzyme
amylaza, esteraza, pectinaza, proteaza. Trong ch c cha nhiu enzyme oxy
ho kh nh polyphenoloxydaza. Trong rau qu cng cha nhiu loi enzyme
khc nhau.
Thu enzyme t thc vt cng khng kinh t v mun c nhiu thc vt
th phi trng nn phi c t v chi ph tn km trong vic chm bn.
5.2.3 Vi sinh vt:
So vi ng vt v thc vt th vic thu nhn enzyme t vi sinh vt c
nhiu li th hn.
- T vi sinh vt c th thu nhn c nhiu enzyme khc nhau, trong
c nhng loi khng th thu nhn c t ng vt hoc thc vt.
- Bng cch thay i iu kin nui cy hoc dng tc nhn iu chnh
ngi ta c th iu khin qu trnh sinh tng hp enzyme ca vi sinh vt theo
mun.
- Vi sinh vt c kh nng sinh sn v pht trin nhanh, tng hp enzyme
vi tc cc k ln trong mt thi gian ngn. Chnh v th vic sn xut
enzyme t vi sinh vt t tn thi gian.
- Enzyme thu nhn t vi sinh vt c hot tnh rt mnh, vt xa cc
enzyme t cc ngun sinh vt khc.
- Vi sinh vt c kh nng sinh sn, pht trin v tng hp enzyme trn
cc mi trng dinh dng n gin, d kim v r tin (c th l ph liu ca
cc ngnh sn xut khc nhau). Cho nn vic sn xut enzyme t vi sinh vt
cng rt kinh t.
Aspergillus oryzae
Amylaza c th thu nhn nh Bacillus Subtilis
Bac.Stearothermophilus
-Amylaza
Asp.niger
Asp.oryzae
Asp.awamori
Glucoamylaza
Asp.oryzae, niger
Endomycopsis, awamori
Asp.niger
Pectinaza
Asp. oryzae
Asp.awamori
Asp.oryzae
Proteaza
Bac.Subtilis
Streptomyces griseus
Proteinaza
Asp.awamori, flavus, Candidus
Bac.mesentericus, Bac.Subtilis
Xenlulaza
Trichoderma viride
Asp.terreus
Kh nng sinh enzym ca cc chng vi sinh vt khng ging nhau v
khc nhau ngay c trong cng mt chng. V vy, khi tuyn chn vi sinh vt
phi tin hnh tm kim, phn lp, la chn hng chc, hng trm thm ch n
hng nghn ging vi sinh vt khc nhau mi c c nhng chng vi sinh vt
c hot lc cao trong vic to nhng enzym cn thit.
2. Ci to ging:
nng cao sc sn xut enzym ca vi sinh vt cng nh lm tng cng
hot tnh ca enzym ngi ta phi s dng cc phng php khc nhau ci
to ging, c bit l phng php gy t bin. Bng phng php t bin
nhiu nc thu c nhng chng vi sinh vt c nhng c tnh tt, ch tng
Nhc:
- Kh c gii ho, t ng ho.
- Nng sut thp.
- Tn nhiu lao ng.
- Tn nhiu mt bng.
2. Phng php nui cy chm:
Vi phng php ny vi sinh vt pht trin trong mi trng lng c
sc kh v khuy o lin tc. Ngun cacbon s dng trong phng php ny l
tinh bt, cc cht thi c ng nh r ng v cc nguyn liu khc. Ngun
nit hay dng l cht hu c nh cao ng, dch men bia, dch thu phn, kh
lcNgoi ra trong thnh phn mi trng cng cn b sung thm cc cht
khong. Mi trng cn c thanh trng bng cch sc hi trc tip
to=121125oC trong thi gian 6090 pht i vi mi trng cha nguyn liu
bt, m th v 2030 pht vi cc loi mi trng c cha ng.
Trong qu trnh nui cy chm cn phi sc kh v trng v khuy o
lin tc. Nu c bt phi dng du ph bt. Thi gian t 14 ngy tu chng.
Phi ch iu chnh pH.
u, nhc:
+ u:
- Tit kim din tch
- D c gii ho, t ng ho.
- S dng hp l cht dinh dng, trnh lng ph.
- Thu enzym t tp cht
- m bo v sinh, v trng nn t nhim vi sinh vt l.
+ Nhc:
- Nng enzym thp, cn lm c canh trng trc khi tch
nn gi thcao.
- Tn in do sc kh.
- Nu khng m bo v trng s nhim hng lot.
3. Phng php nui chm 2 bc:
- Bc 1: vi sinh vt nui trong thit b pht trin ti a.
- Bc 2: chuyn sinh khi sang thit b khc sinh tng hp
enzym.
5.3.3 Tch v tinh ch enzym:
Trong thc t enzym c s dng nhiu dng khc nhau. Mt s lnh
vc c th s dng di dng canh trng kh m cht lng sn phm khng
thay i (nh sn xut ru, cng ngh thuc da). a s cc ngnh cng nghip
(ch yu l thc phm v cng nghip nh) i hi phi s dng cc ch phm
enzym sch (gi l ch phm k thut). Cn trong y hc phi s dng cc ch
phm enzym tinh khit. Tuy nhin, cho n nay cha tm c mt phng
1
-6
10 Kat=16,67 nKat (nanoKat)
60
C6H12O6
2NAD+
2NADH2
2CH3COCOOH
2CH3CHOHCOOH
E.
S 6: C ch tng hp axit lactic t bo vi sinh vt
3. K thut ln men:
chun b mi trng ln men c th s dng r ng hoc cc loi
nguyn liu khc. vi khun pht trin c th cn b sung thm vo mi
trng mt s cc cht dinh dng khc.
C th tin hnh ln men bng phng php lin tc hoc gin on. Ln
men nhit 500C v phi n nh mt cch nghim ngt. Gim nhit ln
men s lm cho hot lc ln men yu v lm pht trin cc vi sinh vt l.
Ngc li, tng nhit s lm v hot enzyme v lm chm qu trnh ln men.
Qu trnh sinh tng hp axt lactic thch nghi pH axt yu (6,36,5).
Tuy nhin, khi ln men do tch lu axit nn lm cho nng axit ca mi
trng tng ln. Nu pH qu thp s km hm s to thnh axt lactic v qu
trnh ln men s ngng trc khi tt c ng chuyn thnh axit lactic. Do
trong qu trnh ln men phi lun lun gi pH mc thch hp. Ngi ta
thng dng cc mui canxi (CaCO3) trung ho v to ra canxilactat
(Ca(C3H5O3)2).
4. Thu nhn sn phm:
Sau khi ln men ngi ta tin hnh x l thu hi dung dch canxilactat
v em kt tinh. Tip theo lm nng chy cc tinh th canxilactat v dng
H2SO4 chuyn n thnh CaSO4 v C3H6O3 nhit 60700C:
Ca(C3H5O3)2 + H2SO4 = CaSO4 + 2C3H6O3
Tip theo tin hnh tch dung dch axit ra khi hn hp. Sau phi tch
st v lm trong dung dch bng than hot tnh ri li tm tch tp cht.
CH3COCOOH
CH3CHO + CO2
-2H
HO
CH3COOH -2H CH2COOH -2H CHCOOH +
CH2COOH
CH2COOH
CHCOOH
CHOHCOOH
Axit xuxinic ax.fumaric
ax.malic
2
oxalacetat
CH2COOH
COCOOH
COCOOH
+
CH3COOH
COOH
HOOCCH2CCH2COOH
OH
Axit citric
S 7: C ch tng hp axit citric t bo vi sinh vt
Phng trnh ln men tng qut:
C6H12O6 + 3O2 = 2C6H8O7 + 4H2O
3. K thut ln men:
Nguyn liu dng chun b mi trng c th l r ng, dch thu
phn tinh bt hoc c th l cc loi nguyn liu khc. Trong mi trng ngoi
ng cn cn phi c cc cht cung cp nit, photpho, kali, km v mt s
nguyn t vi lng khc.