Phn II

Cc ng dng ca Cng ngh sinh hc

Nhp mn Cng ngh sinh hc 237

Chng 7

Cc ng dng trong nng nghip

I. M u
y l lnh vc cng ngh sinh hc c nhiu ng gp quan trng.
Cc sn phm cng ngh sinh hc mi trong nng nghip cha ng trin
vng ha hn i vi ngi tiu dng v nng dn. Hin nay, cc ng dng
cng ngh sinh hc trong nng nghip ang tp trung vo cc hng: chn
lc v bin i di truyn cy trng c c cc c im mong mun
(nng sut cao, phm cht tt, thch nghi vi cc iu kin ngoi cnh bt
li...), nui cy m v t bo thc vt nhn nhanh ging cy trng, sn
xut cc khng th n dng phc v chn on cc bnh thc vt v
ng vt, th tinh trong ng nghim v cy chuyn phi vt nui, ci thin
nng sut v cht lng ca ng vt, nui trng thy sn, ch bin thc
phm...
Nhn chung, trong nhng nm qua cng ngh sinh hc c nhng
tc ng rt tch cc trong sn xut nng nghip, to ra mt cuc cch mng
su sc trong lnh vc ging cy trng, vt nui v ch bin thc phm.
Nhiu kt qu nghin cu c ng dng trong sn xut v em li
nhng gi tr kinh t ln lao. Chng hn, nhiu ging cy trng mang gen
khng su, khng bnh, khng cht dit c c a ra th trng nh
bng, ng, khoai ty, la mch, la nc, c chua, c ci ng... Nhiu
loi vt nui c th tinh trong ng nghim v cy chuyn phi, s
dng hormone sinh trng tng nhanh sc ln v sn lng sa tru,
b, k c sn lng thc phm v cc cht ph gia sinh hc...

II. Ci thin v nhn nhanh ging cy trng

Hng nghin cu c tp trung nhiu nht ci thin v nhn
nhanh ging cy trng l nui cy m v t bo thc vt (plant cell and
tissue culture). y l k thut nui cy v trng in vitro cc b phn tch
ri khc nhau ca thc vt. Ngoi mc ch nhn ging v ci thin di

Nhp mn Cng ngh sinh hc 238

truyn ging cy trng, nui cy m v t bo thc vt cn ng gp vo
vic sn xut sinh khi cc sn phm ha sinh, bnh hc thc vt, duy tr v
bo qun cc ngun gen qu him Cc hot ng ny c bao hm trong
thut ng cng ngh sinh hc nng nghip (biotechnology in agriculture).
Lnh vc nhn ging v ci thin ging cy trng c bn hng chnh:
- Nhn ging trong ng nghim (nhn ging v tnh in vitro) bng k
thut nui cy t bo, m v c quan ca thc vt. Vi k thut ny trong
mt thi gian rt ngn c th sn xut mt lng ln cy con ging ht nhau
v gi nguyn kiu di truyn ca cy m ban u.
- Sn xut cy n bi (1n) bng cch nui cy bao phn hoc ht
phn cho php to ra cc dng thun (ng hp t) phc trng ging cy
trng b thoi ha sau mt thi gian di canh tc. Hoc tm kim cc tnh
trng ln d hp t u vit thu c trong qu trnh chn ging.
- Lai v tnh (somatic hybridization) hay cn gi l dung hp t bo
trn (protoplast fusion) gia cc loi xa nhau v quan h h hng m trong
thc t khng th tin hnh bng phng php lai hu tnh, nh m ra
kh nng to ra nhng ging cy hon ton mi.
- ng dng k thut chn dng t bo bin d soma (somaclonal
variation) trong nui cy in vitro to ra cc ging mi chng chu cc
bnh vi khun, virus v vi nm, chu c cc iu kin canh tc khc
nghit nh hn hn, ngp mn, nng v lnh...

1. Nhn ging v tnh in vitro

Nhn ging in vitro l k thut nhn ging cy trng bng cch s
dng nhiu b phn khc nhau ca thc vt, c kch thc nh v sinh
trng iu kin v trng trong ng nghim hoc trong cc loi bnh nui
cy khc cha mi trng dinh dng nhn to.
Trn quan im ng dng, k thut nhn ging in vitro c ng dng
nhm phc v cc mc ch sau:
- Duy tr v nhn nhanh cc kiu gen qu him lm vt liu cho cng
tc to ging.
- Nhn nhanh vi hiu qu kinh t cao cc loi hoa v cy cnh khng
trng bng ht.
- Nhn nhanh v duy tr cc c th u dng tt cung cp ht ging
cc loi rau, cy cnh v cc cy trng khc.

Nhp mn Cng ngh sinh hc 239

 - Nhn nhanh v kinh t cc kiu gen qu ca ging cy ly g trong
lm nghip v gc ghp trong ngh trng cy n qu, cy cnh.
- Nhn nhanh iu kin v trng, cch ly ti nhim kt hp vi lm
sch bnh virus.
- Bo qun cc tp on ging nhn ging v tnh v cc loi cy giao
phn trong ngn hng gen.

2. Sn xut cy n bi in vitro
>

in vitro

:
- .
- .
- (dng thun).
Mt s phng php c s dng to th n bi nh sau:

in vivo

in vivo

Nhp mn Cng ngh sinh hc 240

 : s

. Nhn chung, cc k thut ny cho hiu

sut to cy n bi thp.

2.2. Phng php to th n bi in vitro

in vitro

, Poaceae,
Ranunculaceae
.

(Hnh 7.1). K thut dung hp protoplast cho php khc phc c hin
tng bt th thng xy ra khi lai khc loi (lai xa) m rng ngun gen,
to ra cc ging cy trng mi mang cc c tnh di truyn u vit.

[Lycopersicum esculentum

258.

Nhp mn Cng ngh sinh hc 241

 A B

Hnh 7.1. Dung hp protoplast. A: cc protoplast. B: hai protoplast dung hp

trong mt cp. C: cc protoplast c th dung hp trong th 3 (bn phi nh) hoc
nhiu hn, c khi ti 6 protoplast.

3.1. Dung hp protoplast bng ha cht

Phng php ny dng NaNO3 hoc polyethylene glycol (PEG)
kch thch s dung hp ca hai protoplast.

3.2. protoplast (electrofusion)

trong n

Nhp mn Cng ngh sinh hc 242

 -

4. Chn dng bin d soma

in
vitro :
-
- - ...
-

- (overproduction)
...
-
(genetic markers)...

Ging ng bt dc c

Nui cy callus

Chn lc trn mi trng cha c t ca

nm bnh r st (Helminthosporium maydis)

Dng callus khng H. maydis

Ti sinh cy

Kim tra tnh khng H. maydis

Hnh 7.2. S c Helminthosporium maydis ng

Nhp mn Cng ngh sinh hc 243

 in
vitro

) , khoai
ty).

-
- ). Tuy nhin, t

5. Chuyn gen vo cy trng

hn

protoplast.

Nhp mn Cng ngh sinh hc 244

 Stt Loi Phng php chuyn gen Th nghim trn
ng rung
1 Chui Bn gen/Agrobacterium -
2 La mch Bn gen Khng vi rus
3 u ty Bn gen -
4 Canola Bn gen/Agrobacterium Chng chu cht dit c,
iu khin s th phn
5 Sn Bn gen/Agrobacterium -
6 Ng Bn gen/Agrobacterium Khng cn trng, chng
chu cht dit c
7 Bng Bn gen/Agrobacterium Khng cn trng, chng
chu cht dit c
8 u Bn gen/Agrobacterium Khng virus
9 u phng Bn gen/Agrobacterium Khng virus
10 Bch dng Bn gen/Agrobacterium Chng chu cht dit c
11 Khoai ty Agrobacterium Khng cn trng, khng
virus, chng chu cht
dit c
12 La Bn gen/Agrobacterium Chng chu cht dit c
13 u tng Bn gen/Agrobacterium Chng chu cht dit c
14 B Bn gen/Agrobacterium Khng virus
15 C ci ng Agrobacterium Chng chu cht dit c
16 Ma Bn gen -
17 Hng dng Bn gen -
18 C chua Agrobacterium Qu chn mun, khng
virus
19 La m Bn gen -

Nhp mn Cng ngh sinh hc 245

 A B

C D

Hnh 7.3. Mt s cy trng chuyn gen. A: ng khng cn trng. B: la mch

khng virus. C: c chua cho qu chn mun . D: khoai ty chng chu cht dit c.

).
Agrobacterium
.

Nhp mn Cng ngh sinh hc 246

 .

bar gus

bar
.

mt cy m h
.

Agrobacterium
c Agrobacterium
Agrobacterium
gus

Nhp mn Cng ngh sinh hc 247

 - -
(Southern b

Phaseolus

Agrobacterium

gusA, bar

gus
gus .

5.6. Cy bng
Agrobacterium
tumefaciens

soma

Nhp mn Cng ngh sinh hc 248

5.7. C nh m
Qu trnh c nh m din ra r ca cc loi cy h u nh mt s
loi vi khun cng sinh c kh nng hp th nitrogen ca khng kh v to
ra cht m cho cy. Vi khun chnh tham gia qu trnh c nh m l
Rhizobium. S c mt ca n dn n hnh thnh cc nt sn trn r cy
nhim khun. Cc vi khun sng trong nt sn s thc hin qu trnh c nh
m. Phng trnh tng qut c dng sau:

Nitrogenase
N2 + 10H+ + 8e- + 16ATP 2NH4+ + 16ADP + 16Pi + H2

Rhizobium Plasmid
RE

Phn lp gen nif M vng DNA ca plasmid

DNA ligase

La Protoplast Bin np

Protoplast mang
plasmid c gen nif

Callus

Ti sinh cy
mang gen nif

Hnh 7.4. M hnh bin np gen nif

tng mc cung cp m t nhin cho cy trng, ngi ta chn

gii php chuyn gen nif (nitrogen fixation-gen m ha enzyme nitrogenase)
vo c th thc vt v vi sinh vt sng t do hoc sng cng sinh vi thc
vt (Hnh 7.4). V trong t nhin, gia cc c th vi sinh vt vn xy ra s

Nhp mn Cng ngh sinh hc 249

trao i thng tin di truyn, nh vy v mt k thut vic chuyn gen nif vo
c th vi sinh vt s d dng hn. Vn cn li l lm th no gen nif
hot ng sau khi c chuyn vo c th vi sinh vt ch. Tng t, vic
chuyn gen nif vo t bo thc vt c th c thc hin thng qua viral
vector hoc vi khun Agrobacterium tumefaciens. Tuy nhin, y l mt
cng vic kh khn v c l trong mt tng lai gn vn ny vn cha
gii quyt c.

III. Chn nui v th y

1. K thut cy chuyn phi
K thut c ng dng kh rng ri hin nay l cy chuyn hp t
b. Nguyn l ca k thut ny l gy rng trng b ci c cc c im
m ngnh chn nui cn n v cho th tinh vi tinh trng ca b c cng
mang nhng c im mong mun. Cc hp t hay phi thu nhn bng cch
ra d con. ng lnh phi v bo qun chng trong nitrogen lng
-179oC, phi iu kin ny c vn chuyn d dng n ni cn thit.
Sau , phi c cy vo b ci khc mang thai h. B con pht trin t
cc phi ny s ra i trong mi trng sng ca n v khng gp phi
nhng bt li v mt mi trng nh i vi cc gia sc nhp ni.
K thut cy phi cn cho php to ra gia sc sinh i hoc sinh ba
(hoc nhiu hn na) hon ton ging nhau v mt di truyn. Nguyn tc ca
k thut ny l chia non bo th tinh ca gia sc thnh hai hoc ba phn
(hoc hn na) ging nhau. Sau , cy ngc li vo con ci cha h.
Mt khc, k thut cy chuyn phi cn gip xc nh gii tnh ca
vt nui, vic ny em li hiu qu kinh t cao. Chng hn, ngi ta ch
quan tm ly phi b c to b ly tht, trong khi phi b ci c dng
to b ly sa, hoc bo v nhng ging b u vit. Nguyn tc ca
k thut ny da trn c s phn chia tinh dch thnh hai nhm: nhm cha
nhim sc th X to ra b ci, v nhm cha nhim sc th Y to ra b c.
Ngi ta ch cn chn mt trong hai nhm th tinh trong ng nghim ri
sau chuyn phi c to ra vo b m.

2. To ch phm phng trnh bnh cho ng vt

Cng ngh sinh hc trong nhng nm gn y c nhng ng gp
khng nh, ch yu l to ra cc vaccine th h mi nh p dng cng ngh

Nhp mn Cng ngh sinh hc 250

DNA ti t hp, chng hn nh vaccine phng bnh l mm long mng
(Foot and Mouth Disease Virus-FMDV), bnh Theileriosis (bnh st b
bin ng-East Coast Fever) gia sc, bnh st ln Chu Phi, bnh
Newcastle, bnh cu trng gia cm
Bnh toi g do virus gy ra gia cm, c bit l g. Virus ny thuc
nhm Paramyxovirus, bao gm c virus quai b, thuc h Paramyxoviridae
(h ny bao gm c virus gy bnh ln go). Virus c kh nng lm ngng
kt hng cu, thng qua mt loi protein F ca n. Loi protein ny c kh
nng gy min dch v c dng lm vaccine.
Bnh l mm long mng l loi bnh do virus, ly lan rt nhanh v
gy bnh cho khong 30 loi ng vt mng guc, c bit nguy him i
vi tru, b, ln v cu. Khi c dch, trnh bnh ly lan, phi tiu dit
hng lot gia sc nhim bnh, thc hin cc bin php cch ly nghim ngt.
Ngi ta thnh cng trong vic dng enzyme trypsin x l v protein v
nhn thy mt trong s cc protein v ca n c th to c min dch,
kch thch sn sinh khng th. Nm 1985, cc nh khoa hc xc nh
c gen m ha loi protein ny v to dng n trong E. coli, bin E. coli
thnh nh my sn xut protein ni trn v khi tim vo b, ln th chng
tr nn min dch i vi loi virus gy bnh l mm long mng.

3. Chuyn gen vo ng vt
Mt trong nhng th nghim chuyn gen u tin c tin hnh l
gn gen m ha cho hormone sinh trng ca chut cng v promoter
methallothionein ca chut nht vo plasmid vector, sau vector ny c
tim vo t bo trng ca chut nht th tinh. Kt qu l chut nht
chuyn gen ny ln nhanh hn chut nht bnh thng. Kim tra m chut
nht chuyn gen thy rng promoter methallothionein iu ha s biu hin
gen hormone sinh trng trong gan nhiu hn trong tuyn yn mc d
chut thng th tuyn yn l chnh. iu ny cho thy vic chuyn gen
thot khi s kim tra iu ha ca tuyn yn, dn n vic sn xut mt
lng ln hormone sinh trng.
Ln chuyn gen tng hp hormone sinh trng ca ngi tuy khng
ln hn v kch thc nhng li c lng tht nc nhiu hn, t m hn v
tiu tn thc n t hn t 20-30%. Tuy nhin, cc ng vt chuyn gen d b

Nhp mn Cng ngh sinh hc 251

thp khp, stress v mt kh nng sinh sn cng nh nh hng chuyn
ng.
Nh chng ta bit cystein cn cho s pht trin lng, nhng do cu
thiu kh nng tng hp amino acid ny v do cc vi sinh vt ng rut
ca cu s dng phn ln cystein sn c nn nh hng xu n sn
lng lng cu. khc phc iu ny, ngi ta to dng hai gen vi
khun m ha cho enzyme chuyn ha serine thnh cystein. S n nh ca
cc gen ny trong cu chuyn gen sau gip cu t tng hp c
cystein lm tng s pht trin ca b lng.
Hin nay, cc nh khoa hc cng c nhng thnh cng bc u
trong vic chuyn cc gen ngi vo ln nhm mc ch ly ph tng ca
chng (tim, gan, thn...) cy ghp, thay th cho cc b phn b h hng
ca ngi. Nh vy, cng ngh gen m ra cho y hc mt kh nng to ln
trong lnh vc cy ghp cc b phn cho ngi v mt trin vng gii quyt
vn min dch cy ghp.
Bn cnh vic ch i nhng thnh cng mi ca cng ngh chuyn
gen trc tip cho vt nui, ngi ta cng ang tm kim cc bin php mi
tng nng sut vt nui bng cng ngh DNA ti t hp. Chng hn, mt
loi hormone tng trng ti t hp ca b (recombinant bovine growth
hormone) c chuyn vo E. coli sn xut thnh cng loi hormone
trn, v c th kch thch tng sn lng sa b ln ti 15% m khng cn
phi gia tng thm khu phn thc n hng ngy. Hoc gen sn xut
hormone sinh trng ca g cng c a vo E. coli, v ngi ta
chng minh c rng hormone c sn xut bng k thut trn cho php
tng kh nng sinh trng ca g ln khong 15%.

IV. Ch bin thc phm

y l lnh vc cng ngh sn xut thc phm t nng sn c s tham
gia ca vi sinh vt, cng nh s ng gp ca cng ngh sinh hc hin i
trong vic nng cao hiu sut ca cc quy trnh sn xut ni trn. Thc
phm c to ra bi vi sinh vt rt a dng, t cc sn phm truyn thng
c ngun gc xa xa nh men bnh m, phomt, sa chua, ru vang, ru
ct, n nhng sn phm mi xut hin nh protein nm (mycoprotein)...
Trc y, trong cng nghip thc phm cc nghin cu cng ngh
sinh hc c s dng ch yu hon thin cc quy trnh cng ngh ln

Nhp mn Cng ngh sinh hc 252

men truyn thng. Cn hin nay, cc nghin cu cng ngh sinh hc ch
yu lin quan n vic to ra cc chng mi c nng sut sinh hc cao v
vic p dng chng vo cc cng ngh ln men hin i.
Theo nh gi chung, hin nay mi ch khong 15% thc phm trn
th gii c sn xut bng cc quy trnh cng ngh sinh hc. Do vy, nh
hng ca cng ngh sinh hc hin i trong lnh vc ny thc s cha r
nt. Nguyn nhn c th l do ngnh cng nghip ch bin thc phm hin
nay vn mang nng tnh th cng, cng ngh cha hin i, ngnh cng
nghip ch bin thc phm c quy m to ln nhng li c li nhun nh, do
vy kh c kh nng ti u t.
Tuy vy, trong tng lai khi cng ngh ch bin thc phm truyn
thng khng p ng c nhu cu ngy cng gia tng ca con ngi, th
nh hng v vai tr ca cng ngh sinh hc hin i s gia tng rt mnh.

1. Sn xut sa
Cc sn phm sa quen thuc i vi chng ta u c to ra trong
qu trnh ln men ca mt s nhm vi khun nh Lactobacillius,
Streptococcus, Leuconostos... Trc kia ngi ta thng s dng nhng
nhm vi khun t nhin c mt trong sa ln men, do vy qu trnh ln
men ni chung kh kim sot v hiu sut khng cao. Ngy nay, nh vic
to ra c cc ging, chng vi khun vi cc tnh cht xc nh, ngi ta
c kh nng iu khin c qu trnh ln men ni trn mt cch c nh
hng. Cc sn phm ch yu t sa l phomt, sa chua, b, kem sa...

1.1. Sn xut sa chua

Trong sa c sn mt h vi sinh vt phong ph, l vi khun lactic
(gy ln men lactic), vi khun acetic (ln men acetic), vi khun ng rut,
vi khun gy thi (phn gii cht hu c thnh cc mi thi), nm men, nm
mc...
Trong sn xut sa chua bng ln men lactic, vi khun dng ln
men chnh l mt s chng thuc Lactobacterium bulgaricum... c sn trong
sa. Trong cng nghip sn xut sa chua, ngi ta tin hnh thanh trng
sa ri cy vi khun lactic vo.
Quy trnh sn xut sa chua theo quy m cng nghip nh sau:

Nhp mn Cng ngh sinh hc 253

1.1.1. Nguyn liu
Nguyn liu duy nht l sa (c th b sung thm ng). Sa dng
trong sn xut sa chua gm nhng loi sau:
- Sa ti nguyn hay sa tch bo.
- Sa kh nguyn hay sa sy phun khng bo.
- Sa c nguyn hay sa c khng ng.
Cc loi sa trn trc khi a vo s dng cn phi x l qua cc
khu:
- iu chnh hm lng lipid thch hp thng t 3,2-6%.
- Nu trn b mt sa xut hin vng sa th tin hnh qu trnh ng
ha tr thnh mt th ng nht.
- Tit trng sa 85-90oC trong 15-20 pht. ngui sa n nhit
cn thit cho s ln men cy ging vo, thng t 40-50oC.

1.1.2. Ging
Cc chng lactic thun khit thng c dng sn xut l:
Streptococcus thermophilus, Lactobacterium bulgaricum, Lactobacterium
acidophilum, Lactobacillus delbrueckibulgaricus. Thng hai chng u
c cng s dng vi s lng bng nhau. Nu Streptococcus
thermophilus c s lng nhiu hn, lc sa chua thu c s qu chua
v km mn.

1.1.3. Ln men to sa chua

Quy trnh sn xut sa chua qua hai giai on: giai on ng t sa
v giai on gi chn sa chua.
- Giai on ng t sa
Sa sau khi tit trng c cy ging vo v khuy u ln men
bng phng php b hoc chai. giai on ny qu trnh ln men c
tin hnh 29-35oC. Qu trnh ln men lactic xy ra mnh m, casein sa b
kt ta im pH ng in, sa c ng t v t chua cn thit.
Thng cn c vo cc ng ca sa v chua kt thc qu trnh ln
men. Cc ng phi c, ng nht, khng c hin tng nc-sa tch ri

Nhp mn Cng ngh sinh hc 254

nhau ra. chua phi t 60-80oT. Thi gian ln men (thng 8-12 gi) c
th thay i khc nhau ty thuc vo chng vi khun v nhit ln men.
Khi sa c chua cn thit th kt thc qu trnh ln men bng phng
php lm lnh nhanh (6-8oC) trnh tnh trng chua tng v nc s
tch ra khi sa.
- Giai on gi chn sa chua
Trong giai on ny qu trnh ln men lactic vn cn xy ra mc
yu, sa tip tc c ng t. Lc ny lipid trong sa tr nn rn, nc t
do lin kt vi protein lm sa ng c thm. Giai on ny cn xy ra qu
trnh to hng cho sa lm cho sa c mi rt c trng bng cch b sung
vaniline, hng v tri cy t nhin (du, cam, da...) hay hng tng hp,
mu thc phm. Thi gian ca giai on ny t 12-14 gi.

1.2. Sn xut phomt

Phomt l sn phm ln men c ch bin t sa (sa b, sa d...)
vi s tham gia ca mt s nhm vi sinh vt. y l mt thc phm c gi
tr dinh dng cao, bo qun c lu. Trong phomt cha 20% protein
(di dng peptone, amino acid), 30% lipid, cc mui khong, vitamin...
Phomt c mi v thm ngon, kch thch qu trnh tit dch tiu ha,
lm tng kh nng ng ha thc n cho c th. Quy trnh sn xut phomt
c tin hnh qua bn giai on. Cng nh sn xut sa chua, nguyn liu
chnh y l sa.
- Giai on lm ng sa
T nguyn liu sa ban u, sa c lm ng li. V mt ha l
l giai on lm kt ta: nhng micelle ca casein dnh li vi nhau hnh
thnh mt gel c, rn cha huyt thanh sa bn trong. Sau khi c kh
trng 85-90oC trong 15-20 pht, c x l bng rennin vi mt lng
nh va , sau mt thi gian nht nh sa s ng li thnh mt khi nhy
ph gelatin, mm do, khng thm nc.
lm ng t sa, ngoi vai tr ca rennin ngi ta cn s dng vi
khun lactic. Vi khun lactic c to iu kin hot ng 30oC. Khi tin
hnh lm ng t sa, ngi ta cy vi khun lactic vo mi trng sa. Qu
trnh ln men lactic c tin hnh, chuyn ng lactose ca sa thnh
lactic acid. Kt qu l qu trnh ny cng gy ra s ng t sa nh trnh

Nhp mn Cng ngh sinh hc 255

by trn (cc si micelle ca casein sa li kt thnh cc rn cha huyt
thanh sa bn trong). Khi qu trnh ln men lactic xy ra, pH mi trng
s gim, pH hng v mi trng acid l pH thch hp cho hot ng ca
enzyme ng t sa. iu ny dn n s ng t sa cng xy ra nhanh
hn. giai on ny, di tc dng ca rennin, casein v paracasein ca
sa s b phn gii to peptone v amino acid (tyrosine, tryptophan...) tp
trung li trong cc ng. Huyt thanh sa bn ngoi cc ng tn ti
dng dung dch.
- Giai on kh nc
Giai on ny p cc sa tch huyt thanh ra khi cc ng sa.
Qu trnh ny xy ra 35-50oC trong 20-24 gi. Trong thi gian ny qu
trnh ln men lactic vn tip tc. Phomt lc ny c thnh phn ch yu l
casein v lipid.
- Giai on mui phomt
Ngay sau khi p cc ng tch huyt thanh, phomt c ngm vo
b nc mui NaCl nng 24% trong vi ngy tng v mn cho
phomt, to s ng nht v thnh phn cho khi phomt v ngn chn s
pht trin ca vi sinh vt c hi, ch yu l trc khun ng rut. Kt qu
ca qu trnh ngm mui l cc cht b mt ca khi phomt nh ng,
mui khong s khuch tn ra ngoi, ngc li NaCl t ngoi dung dch
ngm s thm vo bn trong khi phomt. Mui n thm vo lp b mt ca
phomt, to lp bo v chng li s xm nhp ca vi sinh vt c hi.
- Giai on chn pho mt
Phomt c a vo hm lm chn 50-57oC c m l 80-90%.
Qu trnh lm chn phomt ko di kh lu t vi thng n hng nm, bao
gm nhiu khu chuyn ha ha sinh phc tp, c nhiu nhm vi sinh vt
khc nhau tham gia cng vi men ng t sa. Hot ng ca vi khun
lactic v rennin c mi lin h mt thit vi nhau dn n lm tng cht
lng ca phomt trong giai on ny.
Sau khi ch bin xong, trc khi c ct ra v ng gi bng giy
nhm, oliofilm, rilsan, phomt c kh trng bng cch chiu tia t ngoi.
Vic ng gi c tin hnh trong iu kin v trng, cui cng thu c
thnh phm.

Nhp mn Cng ngh sinh hc 256

2. Ch bin tinh bt
Sn phm truyn thng t bt m rt a dng v c to ra trong
nhng quy trnh sn xut khc nhau. Tuy nhin, chng c mt im chung
l s dng nm men bnh m Sac. cerevisiae ln men. Cho n nhng
nm 1950, tinh bt ch yu c thy phn bng cng ngh s dng acid.
Vo nhng nm 1960, bng cng ngh thy phn acid kt hp vi x l
enzyme v sau ny ch yu bng enzyme.
Thng ngi ta thy phn tinh bt bng -amylase v
amyloglycosidase, l hai enzyme c tc thy phn cao, khng gy
nhim mi trng v to ra sn phm c h s ng lng dextrose DE
(dextrose equivalent) cao. H s DE phn nh mc thy phn tinh bt
thnh ng glucose (dch ng glucose nguyn cht c DE = 100, cn
dch tinh bt c DE = 0). Hin nay, ngi ta to c mt s chng vi
khun, trong c Bacillus licheniformis v Bac. amyloliquefaciens, c kh
nng tng hp -amylase chu nhit hot ng c nhit ti 100oC.
iu ny cho php qu trnh h ha thc hin c trit , to iu kin
cho giai on ng ha tip theo, to ra dch ng c DE gn ti 100.
Thng thng, ngi ta tin hnh ng ha bng glucoamylase nhn c
t nm Aspergillus niger.
Dch c hm lng glucose cao c s dng vo nhiu mc ch
khc nhau, trong ch yu l lm nguyn liu ln men. Tuy nhin, gn
y mi xut hin hng sn xut sir fructose-glucose t dch glucose.
Hin nay, sn phm ny ang c s dng rng ri trong ngnh ch bin
thc phm cc nc pht trin thay th ng saccharose truyn thng
t ma v c ci ng. Thng thng, sir fructose-glucose c to ra t
glucose ca dch thy phn tinh bt c DE gn 100, thng qua qu trnh kh
ion v ng phn ha dch glucose bi glucose isomerase do Streptomyces,
Bac. coagulans v Actinoplanes missouriensis to ra. Trong , qua mt ln
x l ngi ta nhn c sir cha khong 51% glucose, 42% fructose v
7% oliogosaccharide. Nu x l tip s c th nhn c sir cha ti 90%
fructose.

3. Sn xut nc ung ln men

Cc loi nc ung ln men c cn ni chung u c sn xut t
nguyn liu cha ng. Trong , qu trnh to ethanol bng ln men ca
cc chng nm men thuc ging Saccharomyces l qu trnh chung, cn

Nhp mn Cng ngh sinh hc 257

tng loi nc ung ln men c cn li c quy trnh sn xut ring ca
mnh.

3.1. Sn xut bia

Bia l nc gii kht cha nc, CO2, ru, cc cht chit xut,
carbohydrate... C s khoa hc ca ln men sn xut bia l s trao i cht
trong qu trnh sinh trng, pht trin ca mt s loi vi sinh vt trong iu
kin ym kh.
Qu trnh sn xut bia ch yu da vo s ln men ng ca mm
i mch (hin nay c th thay bng ng, go, k...). Bia sn xut t mm
i mch bao gm cng on ch yu l 67oC enzyme t nhin trong
mm thy phn tinh bt ca ht i mch, b sung hoa bia v dch thy
phn, dch ln men bia, cui cng tch men bia v gi bia ti mt thi
gian bia chn. Bia hi l loi bia s dng ngay khng qua kh trng. Bia
chai v bia lon bo qun c lu v qua kh trng 55-60oC hoc qua
lc, ng chai hoc ng hp bng cc thao tc v trng.
Cc chng nm men c s dng ch yu trong sn xut bia l Sac.
cerevisiae v Sac. calsbergensis. Trong Sac. cerevisiae c s dng
cho c ln men b mt v ln men chm sn xut cc loi nc gii kht
ln men. Cn Sac. calsbergensis dng sn xut bia nh.
Qu trnh ln men bia gm 2 giai on: ln men chnh v ln men ph.

3.1.1. Ln men chnh (ch yu dng ln men chm)

giai on ny t bo nm men sinh trng mnh, mt lng ln c
cht c chuyn thnh ethanol, CO2 v H2O. Thi gian ln men khong 6-
10 ngy. Nhit yu cu t 28-30oC, pH mi trng 5,3-6. Trong giai
on ny c hin tng to bt do kh CO2 to ra lc u tan trong dung
dch, sau vi liu lng cao s tch ra to thnh ti kh, to c s cho s to
bt. Kt thc giai on ln men chnh to ra c sn phm gi l bia non.
Bia non c c im cn c, c mi v c trng ca bia nhng cha thch
hp cho vic gii kht.

3.1.2. Ln men ph
Khc vi ln men chnh xy ra trong cc thit b ln men h, ln men
ph thc hin trong cc bnh kn nhit 0-5oC. giai on ny qu trnh
ln men din ra chm, dch ln men c lng ng v bo ha CO2. Giai

Nhp mn Cng ngh sinh hc 258

on ny cn gi l qu trnh chn bia. Thi gian ln men ph ko di t
vi tun n vi thng. Kt thc giai on ny thu c sn phm dng
dung dch bo ha CO2, c hng v thm ngon, d chu nh nhng qu
trnh chuyn ha xy ra nhit thp. Trong giai on ln men ph
nhit thp, xy ra qu trnh ng t nha hoa hublon, cc hp cht tanin-
protein, t bo men lng xung y bnh v tip tc ln men t t. Kt qu
lm bia trong dn. Nh vy, qu trnh ly tm v lc bia thnh phm sau ny
s d dng hn.
Mun lm trong bia thng ngi ta s dng phng php lc (dng
cht bt tr lc diatomite). C th dng phng php ly tm nhng sau khi
ly tm cn lc bia qua lc ng, lc khung bn khng cn cht tr lc th bia
mi tht s trong sut. Trong qu trnh sn xut bia ngi ta s dng cc ti
cht do hay cao su thu hi CO2 thot ra, sau em bm vo cc bnh
chu p lc cao. S kh ny c dng np li vo trong bia, bia thnh
phm c CO2.
tng bn ca bia trong thi gian di ngi ta kh trng 60-70oC
trong 30 pht ngn chn s pht trin ca vi sinh vt v cc qu trnh
bin i trong bia. Bia thnh phm c mu vng, trong sut, hng v thm
ngon, bt nhiu, c v ng cht nh c trng. Bia thnh phm c bo
qun trong kho lnh 0-10oC.

3.2. Sn xut ru vang

3.2.1
Trong

.
- .

(nho c
).

Nhp mn Cng ngh sinh hc 259

 - ).
.
- .

: (1) S ,q

,n
. (2) S
2
O2

2
-

-
( ,
du da...
+

.
+

1...

Nhp mn Cng ngh sinh hc 260

 65-70o .

- . :
+

men thng c s dng: Sac. ellipsoideus, Sac. cerevisiae, Sac.

oviformis
+

.
- . :
+

.
+ .

20-30o n.
+ .

O2 .
- . C ba giai on:
.
-
.
. Sau giai on mt, t

Nhp mn Cng ngh sinh hc 261

 -

-
-

3.3. Sn xut ru trng (cn)

Quy trnh sn xut ru trng bng phng php ln men ru bi
nm men c thc hin qua cc bc sau: ch bin nguyn liu thnh dch
ng, ln men bin ng thnh ru, chng ct v tinh ch ethanol.

3.3.1. Ch bin nguyn liu thnh dch ng

- Nguyn liu. c dng ph bin hin nay l loi nguyn liu c
sn ng (r ng) v nguyn liu c tinh bt (bp, sn, khoai lang, bt
go...).
- Cc phng php ng ha. C ba phng php ph bin nht l
ng ha bng bnh men, maltase v myco-malt.
+ Phng php bnh men. L phng php sn xut ru th cng.
Nm mc (c h enzyme amylase phn gii tinh bt) c nui cy v pht
trin trn mi trng c tinh bt sng. Tinh bt sau khi c chuyn sang
dng d tan (dng nhit cao 130-140oC) m khng nht thit phi h ha
thnh dung dch, th amylase ca nm mc s d dng phn hy tinh bt.
Nhc im ca phng php ny l tinh bt khng dng dung dch nn
hn ch tc dng ca amylase v vic s dng bnh men d gy nhim

Nhp mn Cng ngh sinh hc 262

khun, nn phng php ny to ra nhiu sn phm trung gian v hiu sut
tng thu hi thp.
+ Phng php maltase. S dng ch yu l cc enzyme v -
amylase ca ht i mch hoc tiu mch ny mm (malt) chuyn ha
tinh bt ( h ha) thnh ng ln men. u im ca phng php ny
l: thi gian h ha tinh bt ngn, cht lng ru khng b nh hng m
thng to ra nhng hng v c trng d chu, t b nhim khun. Tuy
nhin, phng php ny cng c mt vi nhc im: hiu sut ng ha
khng cao v khng trit v phc h amylase trong mm thc khng hon
chnh, t l malt s dng so vi hm lng tinh bt c trong nguyn liu
tng i cao (8-20%), gi thnh sn phm cao.
+ Phng php myco-malt. y l phng php c dng ph bin
hn c. Qu trnh ng ha y dng enzyme ca vi sinh vt (ch yu l
ca nm mc). a s nm mc c s dng thuc chi Aspergillus nh:
Asper. niger, Asper. oryzae, Asper. flavus, Asper. awamori. Cc chng nm
mc c s dng ng ha tinh bt c nui cy theo hai phng
php: nui cy b mt (thng dng cho Asper. awamori v Asper. oryzae)
v nui cy chm trong h ln men (thng dng cho cc chng Asper. niger
v Asper. batatae).
Thi gian thc hin qu trnh ng ha di hay ngn ty thuc vo
chng ging nm mc v nguyn liu c s dng, thng thng t 24-40
gi. Cht lng ca qu trnh ng ha c nh gi theo hot lc ca
amylase, dextrinase, gluco-amylase. Kt qu, qu trnh ng ha cho cc
sn phm gm hn hp dextrin, maltose, glucose. Trong , glucose chim
t l cao nht.

3.3.2. Ln men bin ng thnh ru

y l giai on quan trng nht trong sn xut ru, quyt nh cht
lng sn phm to thnh. Sau khi dch ng ha c x l, ngi ta
b sung thm mt s thnh phn cung cp thm vitamin v amino acid
nh mui ammonium, mui phosphate, dch thy phn nm men. Mi
trng c thnh phn nh trn c th s dng ln men.
- Ging. Ch yu l cc chng ca nm men Sac. cerevisiae. Cc
chng nm men dng trong sn xut phi c nhng c im c bn sau:

Nhp mn Cng ngh sinh hc 263

 + C y c im c trng ca nm men.
+ Tc pht trin mnh, hot lc ln men cao.
+ Ln men c nhiu loi ng khc nhau v t c tc ln
men nhanh.
+ Chu c cn cao t 10-12%.
+ Thch nghi c vi nhng iu kin khng thun li ca mi
trng, c bit l i vi cht st trng.
- Qu trnh ln men. Mi trng ln men sau khi c kh trng,
kim tra ng t 90-120 g/L v pH 4,5-4,8 th c th cy ging vo.
Thi gian ln men t 65-72 gi, trong 10 gi u c sc kh nm men
sinh si ny n, sau cho ln men tnh (ym kh). Qu trnh ln men ru
xy ra nh sau: ng v cc cht dinh dng ca mi trng ln men c
hp th vo trong t bo nm men qua mng t bo v tham gia vo qu
trnh trao i cht, ru ethanol v CO2 to thnh lin thot ra khi t bo,
ru ethanol ha tan mnh trong nc do vy n khuch tn rt nhanh vo
mi trng chung quanh.

3.3.3. Chng ct v tinh ch ethanol

Khi kt thc ln men ru, sau khi loi b t bo nm men, mun
c ru tinh khit cn chng ct dch ln men loi b tp cht. K
thut chng ct ru nh hng rt ln n cht lng ru thu c. ng
thi, t l tp cht v cht lng ru li chu nh hng bi nguyn liu
nui cy. Qu trnh chng ct ru din ra theo cc giai on sau:
- Chng ct dch ln men. Qu trnh ny cho php thu c ru th
bng cch tch cn cng cc cht d bay hi ra khi dch ln men.
- Tinh ch ru. L qu trnh tch cc tp cht ra khi cn th thu
cn tinh khit. Trong cn th ngoi ethanol cn c nhiu tp cht. Da vo
khi lng phn t v kh nng bay hi ngi ta chia lm ba nhm:
+ Tp cht u (c nhit si thp hn ethanol: aldehyde acetic,
ethyl acetate, methyl acetate, methanol) c ly ra giai on u ca qu
trnh tinh ch, c gi l ru u hay cn cng nghip.
+ Tp cht cui (c nhit si cao hn ethanol v kh bay hi, l
ru cao phn t: isoamylic, isobutylic) loi ny t tan trong nc, c
gi l du fusel hay ru tp.

Nhp mn Cng ngh sinh hc 264

 + Tp cht trung gian, ty thuc vo nng ru v tnh cht vt l
ca cc tp cht m n s bay hi cng vi tp cht u hay li vi tp
cht cui. S tp cht ny kh tch khi ethanol khi tinh ch, chng hn:
isobutyrate, ethyl isovalerianate. Sau khi tinh ch, tch ba loi tp cht trn,
ta c ru tinh khit. Hn hp ru-nc l hn hp ng ph (c im si
chung) nn vi phng php chng ct thng thng khng th tinh ch
c ru ethanol c nng ru >95,5% (th tch). V vy, mun c
ethanol tuyt i (>99%) ta phi tinh ch thm (v d: bng phng php
chng luyn di p sut thp p 0,0525 atm) hn hp ru-nc khng
c im si chung v cui cng s thu c ethanol tuyt i.

4. Cc sn phm cha protein

4.1. Thc phm ln men truyn thng giu protein
Bao gm cc sn phm khc nhau giu protein nh phomt, u ph,
nc mm... v cc hnh thc bo qun thc phm nh ln men acid nh
trong sn xut cc loi gi ch, xc xch, c p, c mm... Tuy nhin, cho
n nay cc ng dng ca cng ngh sinh hc hin i trong vn ny ni
chung khng ng k tr ngnh sn xut phomt. Ngc li cc ng dng
ca cng ngh sinh hc li th hin rt r nt trong vic to sinh khi vi sinh
vt v ch bin chng thnh cc dng thc phm khc nhau.

4.2. Protein n bo (single cell protein-SCP)

Protein n bo l mt thut ng c gi theo quy c dng ch
vt cht t bo vi sinh vt c s dng lm thc n cho ngi v ng vt.
Thut ng ny hon ton khng chnh xc, v sn phm c to ra thng
khng phi l mt protein thun khit m l cc t bo c x l thuc
nhiu loi vi sinh vt khc nhau bao gm c vi khun, nm men, nm si v
to.
Sinh khi t bo vi sinh vt lun c hm lng protein cao. Do vy,
trong nhng nm gn y lnh vc cng ngh sinh hc rt ch nghin cu
cng ngh sn xut v kh nng ng dng ca protein vi sinh vt. Qu trnh
sn xut SCP gn lin vi cng ngh ln men, do vy nng sut to protein
ca n rt cao (30-80% khi lng kh). Mt nhc im quan trng ca
SCP l thng cha hm lng nucleic acid cao, c bit l vi khun,

Nhp mn Cng ngh sinh hc 265

nhng cng c nm men v nm si. Nu cc loi SCP ny c s dng
cho ngi th l mt vn , v ngi thiu uricase xc tc cho s oxy ha
uric acid thnh allantoin ha tan hn. n nhiu cc dn xut ca purine s
lm tng hm lng uric acid trong mu, acid ny s kt ta v to thnh
tinh th trong cc khp v ng gp vo vic to nn cc vin si trong
ng niu.

4.2.1. Protein t ngun carbohydrate

Cellulose l cht hu c thng gp nht trn tri t v hng nm
c ti to vi mt khi lng khng l. Cc loi rm r cha ti 30-45%
cellulose. Cellulose cng gp nhiu trong b ma, nc thi cng nghip g,
cng nghip dt v cc cht thi ca cc ngnh cng nghip thc phm. Sn
xut SCP t c cht cellulose l mt hng c nhiu trin vng. Cc vi sinh
vt thch hp cho vic s dng cellulose l x khun a nhit, vi khun t d
c ca cc ng vt nhai li v nhiu loi nm si khc... Trong t nhin, t
gp cellulose thun khit m n thng nm di dng lin kt vi cc
polymer khc nh lignin, pectin, hemicellulose... Lignin l mt polymer
c to nn nh s ngng t ca cc gc ru. Lignin bao quanh cc si
cellulose bng mt mng li ba chiu v do vy ngn cn s phn gii
cellulose nh enzyme. Ring vic lm gim ln ca ht cho php tng
ng k s phn gii cellulose.
n nay, mi ch c sn phm protein t nm (mycoprotein) l protein
vi sinh vt duy nht c cho php s dng lm thc phm cho ngi v gia
sc. Mycoprotein l dng thc phm cha si nm Fusarium graminearum.
Sinh khi c to ra trong qu trnh nui cy nm lin tc trn mi trng
cha glucose v mui ammonium. Sau khi qu trnh ln men kt thc, ngi
ta tin hnh x l nhit i vi sinh khi nm thu c nhm mc ch gim
lng ribonucleic acid ca n. Cui cng tch si nm bng cch lc chn
khng. Quy trnh cng ngh ny c nng sut to sinh khi giu protein cao
hn rt nhiu ln so vi chn nui.

4.2.2. Protein t vi khun lam c nh m v vi to

Vi to (microalgae) l tp hp nhng loi to c kch thc nh b v
c th thch hp c vi vic s dng cc phng php nui cy i vi vi
sinh vt. Vi khun lam (cyanobacteria) trc y c gi l to lam, hay

Nhp mn Cng ngh sinh hc 266

to lam-lc. Trong phn ny, chng ta xp hai loi ny chung vi nhau v
gia chng c nhiu c im sinh l khi nui cy ging nhau. Do , qui
trnh sn xut thu sinh khi ca chng v c bn l khng khc nhau.
Ngay t nhng nm 1940, ngi ta bit mt s vi to c kh nng
trong mt thi gian ngn to ra mt lng sinh khi rt ln cha ti 40-60%
protein khi lng kh (Chlorella sp.) v 50-55% protein khi lng kh
(Scenedesmus). vi khun lam, c bit l chi Spirulina hm lng protein
ln n 60-70% khi lng kh. Hm lng cc amino acid ca nhng
protein vi to v vi khun lam kh cn i gn vi quy nh ca protein
tiu chun. c bit l lng amino acid khng thay th trong protein rt
cao c khi ln n 42%.
n u nhng nm 1960, to lam c nh m cng c bt u
nui trng rng ri trn th gii, bnh to c s dng lm thc phm cho
ngi dn, cn bt to c s dng rng ri lm thc phm b sung cho tr
em suy dinh dng v tng tch sa cho sn ph. Chlorella c nui trng
i tr t thp nin 1950. nui trng quy m cng nghip vi to v vi
khun lam, ngi ta s dng hai h thng c bn l h thng kn v h.
- H thng kn. L mt h ln men, dng nh sng nhn to c cng
cao v c sc kh CO2. u im ca phng php ny l khng ph
thuc kh hu thi tit, iu kin nui cy c kim tra, khng ch mt
cch ch ng. Nhng phng thc ny c gi thnh cao nn kh p dng
rng ri.
- H thng h. L cc b nui s dng nh sng t nhin. Nhc
im ca h ny l cc lp t bo phn y b nui thc t s khng c
tip xc vi nh sng mt tri tin hnh qu trnh quang hp (Hnh 7.5).
Vic thu hoch sinh khi vi khun lam v vi to l cng on rt quan
trng nh hng n gi thnh sn phm. C nhiu phng php thu sinh
khi khc nhau nh: ly tm, vt, lng kt ha hc, lng kt bng in
trng, t lng kt... Trng hp cc vi to nh Chlorella, Scenedesmus...
c kch thc t bo nh th ch yu dng phng php ly tm thu sinh
khi. Vi Spirulina ngi ta s dng phng php lc. Thu hoch sinh khi
cc vi khun lam ny bng mng lc nghing kt hp vi ht chn khng.
Sau khi thu hoch, sinh khi c sy kh bng cc cch nh sy ng kh,
sy chn khng, sy hnh trng... c c thnh phm.

Nhp mn Cng ngh sinh hc 267

 Hnh 7.5. Nui to Spirulina Thi Lan v bt to c ng vin

5. Ch bin rau qu
Rau qu c bo qun lu khng bin cht v trong mt s trng
hp gi tr dinh dng ca chng cn c gia tng nh c x l thng
qua ln men lactic nh vi khun Leuconostoc mesenteroides v
Lactobacillus plantarum.
Ch bin v to ra cc thc phm c gi tr t u tng nh ln men
vi sinh vt c bit t rt lu. Ph bin hn c l cc loi nc chm v
u ph t u tng nh ln Asper. oryzae v Asper. tamari.
sn xut u ph trc ht phi ngm u, sau nghin u
thnh bt v lc qua vi. To kt ta t dch ni trn bng mui Ca hoc
Mg, sau ng thnh bnh u. Trong thi gian , nm si trng pht trin
rt mnh trn b mt bnh u v to hng v c bit cho n.
Hin nay, trong cng nghip sn xut nc qu ngi ta s dng rng
ri cc loi enzyme pectinase, cellulase, hemicellulase, amylase, v
protease. Ch yu x l lm trong nc qu, gim nht, gip qu trnh
lc v n nh ca cht lng ca nc qu p. Trong , qu trnh thy
phn pectin nh pectinase, pectateliase, polygalacturonidase v pectin
esterase c vai tr quan trng bc nht. Chng ct cc lin kt glycoside
trong phn t pectin v do vy, lm tng hiu sut to nc qu cng nh
cht lng ca n.

Ti liu tham kho/c thm

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Nhp mn Cng ngh sinh hc 268

 2. Trn Th Thanh. 2003. Cng ngh vi sinh. NXB Gio dc, H Ni.
3. Nguyn Vn Uyn v Nguyn Tin Thng. 1999. Nhng kin thc c
bn v cng ngh sinh hc. NXB Gio dc, H Ni.
4. Chrispeels MJ and Sadava DE. 2003. Plants, Genes, and Crop
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Hill Publishing Co. Ltd. New Delhi, India.
6. Ratledge C and Kristiansen B. 2002. Basic Biotechnology. Cambridge
University Press, UK.
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Nhp mn Cng ngh sinh hc 269