CNG N TP

K THUT THU HI V HON THIN SN PHM

Phn 1.
1. nh ngha qu trnh thu hi v hon thin sn phm. Mc ch ca qu trnh
thu hi v hon thin sn phm. c im ca sn phm sinh hc? Cc sn
phm i km vi sn phm sinh hc trong qu trnh sinh hc l g? Cc yu t
nh hng n qu trnh thu hi v hon thin sn phm

Cu 1:
-

nh ngha qu trnh thu hi hon thin sn phm

Mc ch ca qa trnh thu hi v hon thin sn phm: Bng cc phng php

khc nhau ngi ta c th thu c nhng cht hay nhng sn phm quan tm ra
khi hn hp cc cht trong dung dch ln men hoc t hn hp cc cht. Sn
phm thu c phi t cc yu cu:
+ Hot tnh cao
+ tinh sch cao
+ Hiu sut thu hi cao
+ chi ph thp

c im ca sn phm sinh hc l d b bin tnh v phn hy, khng chu c

PH, nhit , p sut, a nc, k nc, ha tan trong cc dung mi khc nhau.

Cc sn phm i km vi sn phm sinh hc trong qu trnh lc l nhng sn

phm c kch thc gn bng hoc sp x bng sn phm mnh mong mun.

Cc yu t nh hng ti qu trnh thu hi v hon thin sn phm:

+ nht ca dch ln men

+ Nhit , PH, thi gian trong qu trnh thu hi
+ La chn cng ngh ph hp, xc lp cc bc trong qu trnh tch

+ Nng , kch thc, hnh thi, kh nng kt lng ca t bo vi sinh vt trong canh
trng.
+

Cu 2 .Phng php lc. nh ngha, Nguyn tc, cc yu t nh hng cc phng

php lc v ng dng ca pp lc trong CNSH
Cu 2: Phng php lc:
-

nh ngha:

Trc: Qu trnh phn tch pha rn khi pha lng da vo s khc nhau v kch thc
qua lp vt liu lc. Phn trong i qua mng gi l dch lc(filtrate), phn rn cn li
dnh trn vt liu lc gi l cn lc(filter cake).
Nay: Phn tch cc thnh phn ha tan khi pha lng da vo s khc nhau v kch
thc vt liu lc. Phn trong i qua mng gi l dch lc (filtrate), phn rn cn li
dnh trn vt liu lc gi l cn lc (rententate).
-

Nguyn tc: + Da vo s chnh lch p sut dng vo p sut sau khi lc

+ Da vo kch thc t bo v kch thc l ca vt liu lc

Cc yu t nh hng:
+ p sut chn lch gia 2 bn b mt lc
+ Din tch b mt lc
+ nht ca cht lng
+ Tr lc ca vt liu lc v cn lc

ng dng ca phng php lc trong CNSH: Lc sn phm cn quan tm ra khi

dung dch cc hp cht. c ng dng trong y, dc, nng nghip

Cu 3: Phng php li tm. nh ngha, Nguyn tc, cc yu t nh hng, cc

phng php li tm v ng dng ca pp li tm trong CNSH

Phng php li tm:

+) nh ngha: da vo lc ly tm, lc ly tm l mt lc qun tnh xut hin trn mi
vt nm yn trong h quy chiu quay so vi mt h quy chiu quan tnh.
+) Nguyn tc: roto quay trn vi tc cao, lm cho cc cht bn trong n b nm ra
pha ngoi v da vo hn hp cc cht c t trng khc nhau.
+) Cc yu t nh hng:
-

Tc lng li tm

chnh lch trng lng ring

nht ca mi trng

Kch thc ht lng

Hnh dng ht lng

+) Cc phng php ly tm:

Cc phng php li tm khc bit nhau tc khc nhau hoc theo gradient t trng
Li tm trong cng nghip:
Trong phng th nghim s dng li tm gin on, RCF 5000 500000g cn trong cng
nghip lin tc hay bn lin tc, RCF kh thp.
C 3 loi li tm chnh:
-

Li tm ng: t RCF 13000 17000g

Roto hnh ng di to iu kin cho cht huyn ph i qua. Bm di y v i
ngang qua Rotor. Ht lng b bn ra vch. Cht lng lc trong i ln trn. Qu
trnh lin tc cho ti khi ng li tm y. Qu trnh li tm dng v Ht lng c
nh k ly ra. Phng php ny hay c dng cho cc ht c h s lng thp nh
protein kt ta.

Li tm a: RCF 5000 15000g

Trong qu trnh quay cc ht bn theo a v t li thnh. C vng lng cao c
th gin on hay lin tc ly cht rn ra. C th dng li tm t bo hay xc t
bo. C th ly ra nh k trong qu trnh li tm: khc nhau ch yu nguyn tc
ly ra.

Li tm nm ngang; RCF 1500 500g

Dng cho sinh khi t bo ln lng nhanh v c th ra nh tinh th khng sinh

hay qu trnh loi nc: dng thu sinh khi nm si hay nm men hay x l bn
thi. Roto nm ngang hnh xilanh nn, bn trong lp vt ti. Vt ti v xi lanh
quay cng mt chiu, nhng khc s vng quay. Di tc ng ca lc li tm,
huyn ph phn tch v cc cu t rn lng trn tng ca Roto.
+) ng dng:
-

Tch sinh khi sau ln men

Tch xc t bo thu dch protein sau qu trnh ph v t bo

Kt ta protein

Kt ta phn on
1. Phng php lng v tuyn ni. nh ngha, Nguyn tc, cc yu t nh hng
cc phng php lng v ng dng ca pp lng trong CNSH
Phng

nh

Nguyn tc

Cc yu t nh hng

php
Phng

ngha
L

Khi lng +) Trng lc ca ht y

php

phng

ht

lng

php lng nng

cng lng

ng dng
l

phng

php r tin, chm,

hay

thng dng

t do di kch thc

loi sinh khi t

tc

dng cng ln th

ca trng lng
lc

nhanh

cng

Trong :
d- ng knh ht
- Chnh lch trng
lng ring gia ht
lng v mi trng
xung quanh (kg.m-3-)
g- lc hp dn (9,81
m.s-2)
+) Lc va chm ln ht
lng

bo.
L phng php
hay c s dng
trong sn xut
ung c cn (bia)
hay x l nc thi
(bn hot tnh),
SCP
Trong sn xut bia
dng chng nm

men chm tng

kh nng lng
Trong :
nht ca mi
trng (kg.m-1.s-1)
v Tc lng
(m.s-2)
qu trnh lng xy
ra nhanh:
Chnh lch trng lng
ring ln

nht

ca

mi

trng thp
ng knh ht ln.
Mun tng kh nng
lng c th dng cht
keo t nh: mui nhm
v st, cc cht keo t
polime:
polivinylalcohol

Phng

Trong sn xut bia

php

phng

hay

tuyn

php dng

dng albumin ca

ni

bt kh

trng kt hp

tch cht

vi t bo nm

rn

men. Trong

ra

khi dch

xut

ru

SCP

vang

sn
t

ln men

Acinertobacter
cerificans: protein
c thu trowng
dch ln men c
c c li bng
phng

php

tuyn ni
2. Cc phng php ph v t bo. Nguyn tc, ng dng ca tng pp, cc yu t
nh hng v u nhc im ca tng phng php
Phng php ph v t bo:
Phng php ph v t bo c p dng thu hi cc sn phm sinh hc ni
bo(protein ni bo), c mt s phng php ph v t bo nh sau:
+) Phng php c hc.
Mt s phng php c hc c s dng cho vic ph v t bo l:
Phng php

Nguyn tc

ng dng

Cc yu t nh

ng ha p sut Dch huyn ph t bo i vo

hng
S ln ph v.

cao

qua van vi vn tc khng i

Cu to van

dng plus (xung ) v b ph bi

Tc dng

p lc tc thi. Mi xung van

Nhit

ng v nn huyn ph t bo

p sut

vo vng m bng kim loi

cng ng vai tr nh tng
trong ca bung. Khi van m,
dch t bo c gii phng v
chu trnh mi li bt u

+) Thit b ng T bo p qua l nh, ph mng Ph v t bo M

ha bng tay

t bo.

gan

+) p sut

T bo c a vo l nh vi Vi khun, t bo
mt p sut cao, lc x ph v thc vt
t bo

+) Thit b ng Tng t phng php p sut Dch huyn ph t

ha

Manton

- nhng cho quy m ln hn

bo.

Gaulin
Xay, nghin ct
+) Xay, nghin

Lc c hc ct

Ph v t bo m c

+) ng ha bng Bm cc mnh ln v x nt cc Cc m c, phn ln

dao

mnh vn hn

t bo ng vt v
thc vt

+) Nghin bng cc X thnh t bo bi cc ma st T bo thc vt v vi

cht mi (nhm, micro

khun

ct)
+) Nghin bng bi

Lc nhanh cng vi cc bi thy Dch huyn ph t

tinh ph v t bo

bo

+) Siu m

S dng sng tn s cao ph Dch huyn ph t Loi tip

v thnh v mng t bo bo

Nhit

20kHz/s. C thit b chuyn dao

Nng

ng sng thnh dao ng c

bo

hc

Cc ng dch

huyn ph t
bo v hnh
dng cc

+) Phng php ha hc
Ph v t bo bng cc tc nhn ha hc nh:
-

Khng sinh: Ampicillin, cycloserine i vi t bo ang sinh trng.

Tc nhn chelates (to phc vi kim loi), EDTA (ly i cc ion trong thnh t
bo)

Dung mi hu c: aceton, butanol, chloroform, methanol.

Cht st trng: triton X-100, sodium lauryl sunphate

Kim

Chaotropic agent(cht ph v cu trc bc 3 protein hay ADN): bin tnh nh urea.

*)u im:
*)Nhc im: Thng lm v hot Protein
+) Phng php vt l:
-

T phn: Nm men

*) u im: Gi thnh r, dng enzyme ca chnh t bo ph hy mng t bo,

khng cn s dng thm cht l no khc.
-

p sut thm thu: hay dng cho enzyme Periplasma: Cn bng trong dch ng
20% ri nhanh chng chuyn vo nc 4C.

ng/tan.

*)u im
*) Nhc im: Ch p dng c vi tng trng hp c th (periplasma)
+) Phng php Enzyme:
S dng Enzyme ph v t bo, thnh t bo b thy phn ph v bng thm
thu
Mt s enzyme thy phn thng s dng:
-

Lyzozyme cho Vi khun gram (+)

Lyzozyme v EDTA cho vi khun Gram (-)

Chitinaza cho nm

Proteaza v -glucanaza cho nm men.

*) u im: an ton
*) Nhc im:
- Phng php t tin, ch dng cho quy m nh
C hai phng php ha hc v enzyme ch yu khng lm ph v t bo m c l
hay lm thm thu t bo hn l ph v t bo ra, da trn lin kt chn lc vi cc
cht thnh t bo lm thay i tnh cht mng, do SP c th i qua.

3. Phng php lc mng, phn loi, ng dng, cc yu t nh hng v cc vn

hay gp, cch chn mng lc.
Phng php lc mng:
Mng l vt liu dng phn tch cc cht mt cch chn lc. Mng c th t
polymer, gm, cc si cacbon v cc c cht kim loi xp.
Tin hnh lc mng: Dch lc qua gi l dch thm thu, dch cn li gi l dch c
c. Vic thm thu chn lc l c s ca phng php lc mng.
Cc th d in hnh ca phng php lc mng: siu lc (thu hi protein), thm thu
ngc (loi mui trong nc), bc hi nc qua mng (tch cn).
y l phng php tn t nng lng, c th thay th phng php tch bng nhit
thng thng (phng php bc hi hay chng ct); rt tt cho cc sn phm t chu
nhit v c th tin hnh lin tc tuy nhin phng php ny c th gy ra hin tng
bo trn b mt mng lc nn hay phi sa cha v v sinh thng xuyn.
+) Phn loi:
Mt s loi mng lc
-

Loi porous: c l, chnh l tnh chn lc ca mng. Kch thc ca l c th t

<10 angstroms (1 nm) n 100 microns (10-5 nm). Tuy nhin nu l lc nh s gy
lc cn ln cho qu trnh lc. gii quyt vn ny ngi ta ngh ra loi mng
khng i xng.

Loi Non-porous: Loi mng khng c l. Lin kt gia vt liu mng v phn t
cht lc ng vai tr quan trng. Tnh thm thu ca mng i vi hn hp cht
lc t l kh nng ha tan ca phn t cht lc trong vt liu lc v kh nng
khuch tn.

Loi mng khng i xng:

Gm c 2 lp: Lp mng c l quyt nh tnh chn lc ca mng v tc dng lc

Lp dy c tc dng v khng c tnh chn lc vi hn hp lc v
khng gy lc cn cho dng lc.
Theo kch thc l lc th chia lm 4 loi:

Vi lc

Siu lc

Lc nano

Lc thm thu ngc

+) ng dng:
-

Trong cng ngh sinh hc: Thanh trng mi trng (lc v trng); thu t bo; loi
b xc t bo; c c; loi b Pyrogen (c t VSV); x l nc thi; thit b ln
men dng mng; thot hi nc qua mng; thm tch mu cho thn nhn to.

Trong cng ngh sinh hc c in: Trong cng nghip bia (thanh trng bia, bia c
nng cn thp, thu hi bia); Trong sn xut ru vang (tit trng, lm trong v
n nh, ru vang c nng cn thp). S dng phng php lc mng s t
c cc tiu ch: khng lm nh hng n mi v, loi ci protein khng bn
nhit, tannin, hp cht mu, nm men, hp cht phenol b oxi ha.

+) Cc yu t nh hng:
-

Tc dch u vo

Kch thc l lc

Kch thc vt liu lc

nht ca dch lc

p sut qua mng lc

dy ca mng lc

+) Cc vn gp phi trong qu trnh lc

-

Tr lc ca mng thay i (Fouling). Hin tng m cht tan ng li trn b mt

mng v trong cc l lc, lm gim dng chy v thay i tnh cht kh nng phn
tch ca mng.

Hin tng phn cc mng

Hin tng tc mng

Hin tng hp ph

To lp gel.

+) Cch chn mng lc

C th tin hnh chn mng lc theo kch thc l lc, kch thc ca vt liu lc.
Theo kch thc l lc chia ra lm 4 loi:
-

Vi lc: Ch yu dng lc sinh khi vi khun, kch thc mng 0.1 10 m. Do

c th loi c vi sinh vt nn cn gi l lc v khun. Thao tc vi chnh lch
p sut thp. Mng lc c th i xng hay khng. c ng dng rng ri trong
lc v khun cho cng nghip dc, ung, thc phm, c c sinh khi.

Siu lc: Cc phn t cht ha tan c kch thc1.0 100 nm hay 500 500,000
daltons b gi li ph thuc vo molecular weight cutoff ca mng. Molecular
weight cutoff c coi l kch thc phn t ca cht m ti h s gi li 90
95%. Mng siu lc cn c chnh lch p sut 2 10 bar n 25 30 bar. Thng
mng khng i xng. ng dng c c protein v tch protein c kch thc
nh hn.

Lc nano: Loi cc cht c kch thc > 0,1 1 nm v khi lng phn t > 500
1000 daltons. Thng loi mui trong cc sn phm thc phm, ung, sa.
Mng cellulose acetat loi 95% mui ha tr 2 v 40% mui ha tr 1.

Lc thm thu ngc . Da trn nguyn tc lc y l do chnh lch p sut (p

sut dng v p sut thm thu). Loi cc cht c kch thc < 0.1 nm v khi
lng phn t < 500 daltons. Mng cellulose acetat loi 96 99% mui NaCl.
ng dng trong x l nc ung, nc thi, nc cho dc, cho thc phm.
4. Phng php trch li hai pha lng. Nguyn tc, ng dng, h s phn b
Phng php trch ly:
L qu trnh thu hi 1 cht hay 1 nhm cc cht vo pha lng. Dng phn tch,
c c, v tinh sch sn phm. Thng l qu trnh trc qu trnh tinh sch v
in hnh l trc bc sc k.
Qu trnh trch ly l bc u trong sn xut thuc khng sinh sau phn tch t
bo. u im ca qu trnh ny l lm gim ng k th tch ca sn phm.
+) Nguyn tc: Da vo ha tan khc nhau ca cc cht trong cc dung dch
khc nhau.
Phng php trch ly hai pha lng:

+) Nguyn l: Trch ly lng lng l phng php s dng 2 cht lng khng trn
ln vi nhau. Cht mun trch ly c kh nng ha tan khc nhau trong 2 cht lng
ny Cht tan chuyn t cht lng ny sang cht lng khc.
+) ng dng: Cht tan v dung mi c bay hi ging nhau; Cht tan chu nhit
km; Nng cht tan thp Cc sn phm sinh hc v sn phm dc phm
phn ln trch li lng lng.
+) H s phn b:
-

H s phn b (Partition coeficient) K: l s phn chia ca cht tan ti iu kin

cn bng gia hai pha lng.
K = y/x
Trong :
y- nng cht tan trong pha trch ly
x- nng cht tan trong pha raphinat
K- H s phn b.
Mong mun K cng nh cng tt. Nu K nh cn trch ly nhiu v th tch cht
trch ly ln. Nu K 0 th khng th trch ly.
H s phn b ph thuc vo nhiu thng s: kch thc phn t b trch li, pH,
dng dung mi, nhit , nng v Mw ca polime hay mui trong cc pha
5. Tnh tan ca protein. Phng php kt ta protein bng mui v dung mi

Tnh tan ca protein: l kh nng lin kt vi cc phn t nc trong mi trng.

*) Cc yu t nh hng ti tnh tan ca Protein:
-

Cu trc v kch thc phn t: Trong mi trng lng, protein dng cu trc
gim thiu cc lin kt ca cc axit amin k nc vi cc phn t nc v gia tng
cc lin kt ca cc gc c cc v tch in vi phn t nc. Cc lc n nh cu
trc protein: lc lin kt hidro, lc Van der Waal, lc dung mi khin cho xu
hng cc gc k nc hng vo bn trong phn t Protein, cc gc mang cc v
in tch hng ra bn ngoi. Khi kch thc phn t protein ln th tan km.

S tch in protein: in tch ca protein c n hng trc tip ti tnh tan ca

protein. in tch ca protein tng s lm cho tnh tan ca protein tng do lin kt
ca phn t nc lng cc. Cch n gin nht thay i in tch ca protein
l thay i pH. Gi tr pH m ti in tch ca Protein =0 gi l pI. Kh nng
ha tan ca protein nhn chung km nht ti pI. Tuy nhin do s phn b in tch
khng u dn n s to thnh phn t moment lng cc, do vy lm tng tnh
tan v s dch chuyn tan km nht khi gi tr pI.

Cc dung mi s dng hon tan protein: Dung mi nh hng ti tnh tan ca

protein thong qua tnh k nc v lc ion. Ti nhit cao, dung mi c th lm
bin tnh protein. Kh nng lm bin tnh c th trnh c ti nhit thp. Mc
kh nng lm bin tnh protein ca mt s loi dung mi:

Methanol < etanol < propanol < butanol.

Cn vi cc gc alkyl di s lin kt mnh hn n cc nhm khng phn cc ca
protein, lm yu cc lin kt k nc bn trong phn t protein v do vy chng
lm bin tnh protein. Lc ion c hai tc dng tri ngc nhau l ha tan (Salting
in: nng ion tng lm tng tnh tan ca protein; cc ion chn cc in tch v
cho php protein gp np to cu trc fold) v kt ta (Salting out: cc ion cnh
tranh vi nc lin kt cc protein. Khi nng mui cao protein s kt ta.)
protein.
*) Phng php kt ta protein bng mui hoc dung mi.
Cc bc xy ra trong qu trnh kt ta:
-

Khuy ban u

To nhn

Tng kt ta nh khuch tn

Tng kt ta nh chuyn ng ca cht lng.

Phn t protein khng tan nu khng c mui ti pI nhng nu thm t mui tr nn

tan(euglobulin). Mt s protein li tan trong nc cng nh trong dch c nng
mui cao(albumin). Tng t nguyn tc ny nhng theo chiu ngc li, ngi ta
cng c th tin hnh kt ta protein bng mui hoc bng dung mi.

M hnh Cohn m t qu trnh kt ta khi nng ion cao:

Log S = KsI
Trong :
Ks l hng s kt ta ph thuc vo protein v mui m khng ph thuoscj vo nhit
v pH khi pH >pI
tan gi thit ca protein khi lc ion =0 ch ph thuc vo nhit khi pH b
nht ti pI.
ng dng :
Kt ta l phng php ph bin thu hi protein t hn hp protein; thu hi protein
t dch ngoi bo.
Cch dng:
-

Dng cc mui v c (Sulfat amon, sulfat natri..)

Dng dung mi hu c (axeton, etanol)

Phn II:
A- Phng php tinh ch sn phm:
Cu 1: c im ca qu trnh tch sn phm sinh hc:
-

Nguyn liu ban u:

+ dch ln men ( vi sinh vt), dch huyn ph t bo (ng vt, thc vt ) hoc
mi trng hn hp.
+ Vt liu sinh hc ( mu, t bo)

- Cc tnh cht s dng trong phng php tch: tnh kch thc, trng lng, tnh
ha tan, in tch, tnh k nc hay lien kt c hiu.
- Cht lng sn phm: tinh khit, hot tnh, tp cht, tnh sinh hc, bn.
Cu 2: Nguyn tc thc hin qu trnh tch sn phm sinh hc:
- Cn xc nh mc tiu tinh sch, thu hot tnh hay cht lng sn phm
cui cng.
- Xc nh tnh cht ca protein v mc tinh sch la chn k thut
ph hp.

- Chn phng php phn tch nhanh, hot tnh protein, thu hi, tp cht.
- Gim ti thiu cht b sung, ph tr
- Loi b cc cht c th gy cn tr qu trnh, v d nh protease
- S dng cc k thut tch khc nhau c bc, da vo kch thc, in
tch, tnh k nc, hay lin kt c hiu.
- Gim ti thiu s bc thc hin: trnh gim nng sut, tit kim thi gian.
Cu 3: Cc tnh cht ca protein nh hng n qu trnh tinh sch:
- Tnh bn nhit: Protein khng bn nhit cao, v vy cn thao tc nhanh
nhit thp v chn dung dch m thch hp tch chit.
- Tnh bn pH: chn iu kin trao i ion, sc k i lc v sc k o pha.
- Tnh bn dung mi: la chn iu kin sc k o pha.
- Kt ta bng mui: chn iu kin kt ta, sc k trao i ion, tng tc k
nc.
- Cc tc nhn khc: chn cht b sung, cht m, pH thch hp.
- Nhy vi protease: Cn loi b, hoc dung cht c ch.
- Phn ng vi kim loi nng: b sung them EDTA hay EGTA vo m.
- Tnh oxy ha: cn dung tc nhn kh.
- Trng lng phn t: La chn mi trng lc gel.
- Tnh tch in: La chn iu kin trao i ion.
- Tnh k nc: chn mi trng tng tc k nc.
- Bin i tnh cht: sc k i lc.

B- Cc phng php sc k:
Cu 1: Khi nim v nguyn tc ca cc phng php sc k:
Khi nim ca Mikhail S. Tsvett (1960): sc k l mt phng php tch trong cc cu
t ca mt hn hp c tch trn mt ct hp th t trong mt h thng ang chy.

Khi nim ca IUPAC (1993): Sc k l mt phng php tch trong cc cu t c

tch c phn b gia hai pha, mt trong hai pha l pha tnh ng yn cn pha kia
chuyn ng theo mt hng xc nh.
Thut ng sc k ch mt tp cc k thut phn tch v iu ch cho php
tch bit cc hp phn khc nhau ca mt hn hp.
-Nguyn tc tch bng sc k: da trn s phn b khc nhau ca cc cht tan
gia pha ng (mobile phase) v pha tnh ( stationary phase).
- H s phn tch:
K= Cs / Cm
Trong :

Cs: nng cht tan trong pha tnh

Cm: nng cht tan trong pha ng.

Cu 3: Nguyn tc ca sc k trao i ion:
- S tch xy ra di i lc kh nhau ca cc ion trong dung dch i vi cc
trung tm trao i ion ( nhm cha ion) trn cht rn l pha tnh ( nha trao i
ion).
-Dung mi l nc hoc hn hp nc- dung mi hu c trong mi trng c
in mi tng i ln cc ion c th tn ti t do v bn.
- Protein l mt i phn t mang in tch, c vng tch in thay i theo pH.
pH m ti protein c in tch bng 0 ( trung ha v in) c gi l im
ng in. trn im ng in, protein tch in m v di im ng in,
protein tch in dng. mt pH nht nh, cc protein s c mt in tch
khng ging nhau, do chng c gi nhiu hay t bng tng tc ion trn mt
nha ion cho v vi mt pha di ng cho.
Cu 4: Cc dng sc k trao i ion:
C 2 dng sc k trao i ion, c phn loi da vo kh nng lien kt vi
cc nhm chc tch in tri du: dng sc k trao i anion v dng sc k trao
i cation.
- Dng sc k trao i anion:
+ Mt khung c cc nhm chc tch in dng c bao quanh bi cht mang
khng tan v ion i tch in m c gi l nha trao i anion.

+ Nhng Protein tch in m c th th ch cc ion i v c bao quanh bi

nha trao i ion.
+ ion i trong dung dch m ra gii sau trao i vi hn hp cc protein,
v gii phng ra protein mong mun t nha trao i ion.
+ S phn tch c da trn mnh lin kt ca protein vi nha trao i
ion.
- Dng sc k trao i cation:
+ Mt khung c cc nhm chc tch in m c bao quanh bi cht mang
khng tan v ion i tch in dng c gi l nha trao i cation.
+ Nhng protein tch in dng s th ch cc ion i v c bao bi nha
trao i ion.
+ Ion i trong m ra gii trao i vi hn hp cc protein, v gii phng ra
protein mong mun t nha trao i ion.
+ S phn tch da vo mnh lin kt ca protein vi nha trao i ion
Cu 2: Phn bit cc phng php sc k: sc k hp ph, sc k phn b, sc k trao
i ion v sc k ry phn t:
* Ging nhau: u da trn s phn b khc nhau ca cc cht tan gia pha ng, v
pha tnh.
- c s dng tinh sch cc sn phm.
* khc nhau:

Nguyn
tc

Sc k hp
ph

Sc k phn
b

Sc k trao
i ion

Sc k ry
phn t

Da vo i
lc khc
nhau ca
cc cht
hp ph
rn ( pha
tnh)

Da vo
tan khc
nhau ( s
phn b
kh nhau)
ca cc
cht gia
pha tnh
lng
(c gi

S tch xy
ra do i
lc khc
nhau ca
cc ion
trong
dung dch
i vi c
trung tm
trao i

s dng cc
vt
liu
rn, c
xp ln,
c cc l
vi kch
thc nht
nh

ry chn
lc cc cu

- i vi cc
cht hp
ph phn
cc,
i
lc ch
yu
do
tng tc
cc cc.
Do nhm
phn cc
trong
phn t
nh
hng ti
kt qu
tch
mnh hn
cc mch
hydratcar
bon
khng
phn cc

bi cht
mang
rn) v
pha ng

ion
( nhm
cha ion)
trn cht
rn l pha
tnh. Pha
tnh
l
nha trao
i ion.

t ty theo
kch thc
v
hnh
dng phn
t.

- Trong sc
k lng,
pha tnh
thng
phn cc
hn pha
ng, tuy
nhin
cng c
nhiu
trng
hp
ngc li.

- Dung mi
l nc
hoc hn
hp
ncdung mi
hu
c
trong mi
trng c
in
mi
tng i
ln

cc ion
c th tn
ti t do
v bn.

- Cc zeolit
tng hp
c gi tr
rt
ln
trong sc
k ry cc
cht kh.

- phn
cc ca
cc cht
c sc
k cng
nh ca
cc pha
tnh
v
ng c
nh
hng ti
qu trnh
tch.

- Cc cht xp
v c, hu
c, polime
tng hp
tch cc
cht v c
v hu c.
-

Nhm
polime a
nc ( loi
dextran
mng)
c dung
tch cc
cht
c
hot tnh
sinh hc

Cu 5: Mc ch s dng sc k trao i ion trong qu trnh tinh sch tch

sn phm:

Sc k trao i ion c ng dng nh mt bc trong qu trnh tinh sch

protein, c s dng sau khi kt ta protein hoc sau lc gel, nu kt ta bng
mui th cn thm tch trc khi sc k, c th dng trc tip ngay sau khi lc gel.
Ngoi ra c th s dng phng php ny trc sc k i i lc v sc k
tng tc k nc.
Cu 6: Cc bc v thao tc thc hin qu trnh trao i ion:
- Hp ph thun nghch protein- enzyme cn tinh sch ( v cc protein c in
tch gn ging) vo nha trao i ion: protein mang in tch s th ch cho ion
i mang in tch tri du v c hp th vo nha trao i ion.
- Kh hp ph cc protein c hp ph:
+ bng cch thay i pH ca dch ra s dn n thay i ion ha v do
thay i in tch tng ca protein.
+ hoc bng cc tng lc ion v tng nng ion i cnh tranh. Cc protein,
enzmyme no c i lc vi nha trao i ion y nht s c y ra trc tin v
ngc li.
+ Cc protein- enzyme khc nhau s c tch chit ra khi ct theo tng phn
tch chit (phn on chit) khc nhau, trong phn chit enzyme cn thu c
nng cao nht.
- Kh nng xc nh bi nhm chc, trao i mnh hay yu ty theo kh nng
ion ha khi thay i pH.
+ kh nng mnh: ion ha trong di pH rng.
+ nh lng kh nng trao i ion lien kt ion, ty thuc vo s nhm
chc.
Cu 7: Tnh cht cc loi nha trao i ion v phm vi ng dng:
* Carboxymethyl sephadex ( CM sephadex):
- c tnh:
+ kch thc ht: 40- 125 m ( kh)
+ kch thc l: gii hn loi tr 200000.
+ pH = 6- 10.

- Phm vi ng dng: s dng trong sc k trao i cation lin kt yu.

* Diethylaminoethyl Sephadex ( DEAE sephadex):
- c tnh:
+ Kch thc ht: 40- 125 m ( kh)
+ kch thc l: gii hn loi tr 30000 Da.
+ pH = 2-9
+ Cng sut: 3-4 meq/g
- Phm vi ng dng: s dng trong sc k trao i anion lien kt mnh v yu.
* Sulfopropyl Sephadex:
- c tnh:
+ kch thc ht: 40- 125 m ( kh)
+ kch thc l: gii hn loi tr 200000 Da
+ pH= 2- 10.
+ cng sut: 2- 2.6 meq/g.
- Phm vi ng dng: s dng trong sc k trao i cation mnh.
Cu 8: Nguyn tc v bn cht ca phng php sc k lc gel? Mc ch v ng
dng ca phng php?
Tr li:
Bn cht SKLG:
Da vo s khc nhau ca kch thc phn t ca cc hp cht. ngi ta cng gi
l lc gel hay sc k ry.
Pha tnh l dung mi trong cc l gel
Pha ng chnh l dung mi chy qua
Ni cch khc, pha ng v pha tnh cng l 1 dung mi hay hn hp cc dung
mi.
Nguyn tc SKLG:
Hnh v trong slide nh

Hnh v nguyn tc sc k lc gel

(A) Cc hnh trn ln biu din cc phn t c kch thc ln i bn ngoi l gel.
Cc chm nh l cc phn t c kch thc nh nm trong l gel
(B) Hnh sc k .
C th tnh h s phn b Kav i vi cc peak , tng t nh Kf ca sc k lp mng.
Kav = (Ve Vo)/(Vt-Vo)
Kav = 0 i vi protein kch thc ln v K av =1 khi protein c kch thc nh nht. c
th xy dng mi tng quan gia Kav vi kch thc phn t protein Ve = Vt= Vo+Vi
Ve: trong th tch ra
Vo: th tch trng v Vt : th tch tng
Th tch bn trong hat Vi= Vt Vo. Cui cng mt lng th tch ht Vg khng cha
cc phn t
ng dng ca SKLG: SKLG c dng xc nh s phn b ca cc phn t
polyme theo kch thc phn t: c dng lm sch, tch cc polypeptide, protein,
cc hp cht cao phn t khc.
Mc ch ca SKLG:
Loi mui:
- Tch nhm
-

S dng tch mui khi protein

Protein thu c trong th tch trng
S dng Sephadex G-25, Bio-Gel-P6
S dng ct mini (v d Bio-Spin-6)

Phn on:
-

S dng ht gel c kch thc tng ng tch phn t protein mong mun
Th tch mu 1-3% th tch Vt
Khong m pH= 2-10

Kch thc gel ln th tc dng ln nhng kt qu tch khng tt.

Cu 9: c im cc loi gel s dng trong phng php sc k gel? Nguyn tc

la chn Pha ng
Tr li:
-

Cc loi gel c s dng:

Gel mm
Gel cng
Gel na cng

Gel c to thnh t dextran, polyacrylamid hoc polymer thin nhin haytng

hp khc.
a.
Gel mm
L nhng hp cht cao phn t c s lin kt ngang ko ng k. loi ny c t s
VT/VN =3 (t s th tch dung mi bn trong gel v bn ngoi gel). Loi gel mm khi
trng, th tch tng ng k.
Dng tch cc hp cht c khi lng phn t thp.
b.
Gel na cng
c ch to bng phng php trng hp. loi gel ph bin nht l sn phm
ng trng hp ca styrol v divinylbenzen hay sn phm trng hp ca vinylaxetat. H
s th tch ca gel ny l: 0.8 - 1.2.
Khi trng, th tch ca chng tng ln khng nhiu: 1.2 - 1.8 ln
Loi sc k na gel c gi l sc k gel xuyn thu(permeation)
c.
Gel cng
Thng l silicagen hay thy tinh xp vi l xp c kch thc xc nh. Thc
cht nhng vt liu ny khng phi l gel. Loi gel cng c h s th tch khng ln: 0.8
1.1
Loi gel ny c dng trong sc k gel cao p.
Nguyn tc la chn pha ng: cc dung mi trong sc k gel phi ha
tan c cu t trong hn hp, thm t trn b mt gel nhng ko b gel hp ph.
Cu 10: Cc bc v thao tc thc hin sc k lc gel
Tr li:
I/ Chun b gel
a. Trng hp s dng gel kh

1a- Tnh lng gel kh cn thit, da vo h s trng n ca gel v th tch ca c t.

a ton b lng gel tnh ton vo dung dch m c th tchgp 2 ln th tch
gel cui cng.
2a- Khuy u gel bng a thy tinh. cho gel n trong thi gian 1 m, nhi t
thng ho c 3 h trong iu ki n 90oC bng bn n nhi t, khuy u gel lun
dng huyn ph.
(Ch : Khng c dng khuy t , s lm v gel)
3a- lp gel lngxung, sau loi cc ht gel b v v bng cch gn lp c
trn. L p li vi c gn 4-5 ln nu cn thit cho ti khi thylp gel lng xung
phn lp ro rng, ri b sung dung dch m saocho lp gel chim 50% th tch.
( Muc ich cua bc nay la tranh s tc c t sau nay do cac phn ht nho gy
ra).
b. Nu gel c cung cp di dang trng n t trc
1b- Ra gel bng dung dch m bng phu thy tinh Buchner loi b dch lng
trong .
2b- Nu gel c cha cc phn ht nh th cn gn lc nh bc 2 phn a
3b- Ha gel vi dung dch m c t l 50% th tch l dung dch m
II/ Chun b c t
4. Kim tra sch ca c t, kim tra li trong c t khng b hng
5. t gi ln bn, kim tra thng ng ca c t. Chun b c t c th tch ln hn
gp 2 ln so vi th tch gel c.
6. Loi b khng kh trong c t bng cch bm dung dch m vo c t t y (bng
xilanh ho c bm) cho n khi ng p qu li chn 0,5 cm. ng np.
7. a dung dch m vo c t t pha trn bng ng ni cho n khi li ngm tv
gi cho van thot trng thi ng khng kh c y ra khi li trn. t ng
ni vo cc cha dung dch m cho n khi kt thc.
8. Lc u gel chun b v vo c t dc theo a thy tinh vo thnh trong ca
c t . dung dch m
vo ch trng cn li, dc thnh ng sao cho khng lm
xo tr n lp gel. ynp.
9. dung dch m vo bnh cha v t bnh v tr cao hn bm. Ni bnh cha
dung dch m vi bm.

10. Ra bm bng dung dch m v ni u ra ca bm vi u vo ca c t hay

c t m r ng. M van ra ca c t v bt u bm vi tc nh. Bm u cho
n khi chiu cao ca lp gel n nh. (khong 1-4 h).
11. Tt bm, ng van ra ca c t, b c t ph, b sung gel nucn.
12. Li ni c t vi bm, m van thot ca c t v nhc li thao tc bc 10 trong
vng 1 gi n nh chiu cao c t. Thay ng ni thm gel.
13. Kim tra li c t nhi bng cch quan st c s dng n chiu t pha sau c t.
III/ Chyc t
14. Tnh th tch dung dch m
cn thit v b sung 50% th tchcn, lc qua mng
lc 0,22 m. iu chnh pH nu cn v vo bnh cha dung dch m.
15. Lp h thng lc gel vi h thng detector v b ph n ghi. Ni bnh dung dch
m vi bm v ra bm bng dung dch m, khng qua c t.
16. Ni u bm vi c t qua van bm, chy h thng bng dung dch m vi tc
nh sn, thu cc phn on v ghi tc dng tc thi.
IV/ Xc nh th tch tch
17. Xc nh th tch trng cah thng (Vo) bng cch chy vi marker v thu c
th tch ra theo bc 20 n 23
Th tchVothng chim 30-33% th tch i vi gel mm (nh agarose) v chim
36-40% th tch ca khi i vi gel cng (silic)
18. Xc nh th tch tng ca h thng (Vt) bng cch chy vi cht nh du v thu
c th tch ra nh bc 20 n 23.
19. Tnh th tch tch ca c t Vi = Vt- Vo
Th tch Vi l lng cht lng i qua c t t im m protein ln bt u xut hi n
cho n khi nhng ht nh c ra ra. N chnh l th tchl ca lp gel, thng
chim 50-65% th tchlp gel i vi gel mm v 30-50% i vi gel cng.
V/ Chun b thc hi n mu phn tch
20. Ha mu v loi mui trong dung dch m. Lc qua mng lc 0,22m.
- Cn mu t nhi t phng trc khi chy c t trnh khi cn tr ca
nht cao. Thng thng s dng mu c hm lng protein 50 mg/ml-70 mg/ml. Cn
nng 100 mg/ml s nht.

21. M van y, b t bm, cho dch m chy qua c t (2 ln th tch gel). B t

detector v chy nn cho n nh.
22. Chy mu vi m t th tch tng ng loi mui, khng qu th tch tch
c xc nh bc th 19. B t van mu vo v tr load v b t vo du start
chy giy in.
23a. Xc nh th tch ra. Cho dung dch m chy qua h thng vi tc tng
ngv p ng detector khi thu cc phn on. Xy dng sc k v tnh th tch
ra bng cch ly thi gian k t im xut pht cho n khi t peak nhn vi tc
dng chy .
23b. Loi mui. Cho m t th tch dung dch m bng th tch trng tr m t na th
tch mu chy qua dung dch v thu phn on. t bnh thu di c t c th
cha 1,5 ln th tch mu, sau b sung th tch dung dch m bng vi th tch
mu. Thu protein loimui.
24. Ra c t bng dung dch m c cha cht khng khun vi m t lng ln hn
th tch c t. ng van x ca c t v bo qun c t.
Cu 11: Bn cht ca phng php sc k i lc
Tr li:
SKAL l phng php tch hn hp cc cht da trn tng tc sinh hc c trng
gia
-

Antigen v antibody
Enzyme v c cht
Cht nhn v cht gn

SKAL xem nh l phng php sinh hc hin i tinh sch axit nucleic, protein ti
t hp, dch chit t bo.
Bn cht:
-

Sc k hp ph
Hp ph c trng i vi cht gn(ligand)-phi t
Qu trnh thun nghch
Cht gn c c nh vo cht mang ko ha tan(matrix)

Cu 12: Cc loi sc k i lc? C nhng tng tc lin kt sinh hc no? V d

c th

Tr li:
Mnh tm c ti liu khng chnh thng nhng c vit ra cc bn tham kho
Cc loi SKAL:
Da vo cht c s dng trong pha tnh:
-

Sc k i lc lectin.
Sc k i lc boronate.
Sc k i lc protein hay protein G.
Sc k i lc min dch.

Nhng tng tc lin kt sinh hc:

Enzyme c cht, cht km hm, cofactor
Khng th khng nguyn, virus, t bo
Lectin polysaccharide, glycoprotein, cht nhn b mt t bo, t bo
Hormone, vitamin receptor, carrier protein
Glutathione glutathione S transferase hoc GST fusion proteins
Ion kim loi poly(His) fusion proteins, native proteins with histidine, cysteine
and/or tryptophan residues on their surfaces
V d c th:
SKAL lectin l dng sc k i lc m trong lectin c s dng tch cc
thnh phn trong mu. Lectin l protein c th gn kt c hiu vi phn t ng.
ng dng quan trng l tch cc protein da vo cc nhm glycosyl ha.
Lectin l mt loi protein khng c ngun gc min dich c kh nng lin kt
thun nghch, phi ha tr vi carbonhydrate m ko thay i cu trc ca
carbonhydrate c lin kt. lectin gn kt vi nhng t bo glycoprotein hoc
glycolipid b mt.

Cu 13: Cu to, bn cht ca cht gn kt v cht mang (matrix). Tnh cht ca

cc cht mang? Ly v d c th chn cht gn kt tng ng vi nhm chc
lin kt.

Tr li:
Tnh cht ca cht mang:
Tr v mt vt l v ha hc
Khng tan
C nhm ha hc ph hp gn phi t
Phi bn trong sut qu trnh gn mu
C cu trc ng nht i vi tc dng chy.
Cc dang cht mang:

Cellulose
Agarose, cross-linked agarose
Polyacrylamid
Thy tinh xp/ gm xp
V d c th chn cht gn kt tng ng vi nhm chc lin kt:
Ligand(cht gn kt)

Nhm chc nng

Protein, peptide, aminoacid

Amino, cacboxyl, thiol

sugar

Hydroxyl, amino

polynucleotide

Amino, mercurated base

Coenzyme, cofactor, antibiotic, steroid

Amino, cacboxyl, thiol or hydroxyl

Cu 14: Th no l sc k tng tc k nc? Nguyn tc ca phng php?

ng dng ca phng php
Tr li:
SKTTKN (HIC: hydrophobic interaction chromatography)l mt k thut hp ph
protein nh tng tc phi ng ha tr gia cc vng khng phn cc trn b mt protein
v mng li k nc tch protein ra khi cc thnh phn khc.
Tin hnh c trng nng mui cao, ra gii bng cch gim nng mui.
Nguyn tc:
Hnh v trong slide

Cc phn t nc c sp xp trt t quanh mng protein v phi t c nh

Khi protein gn kt vi cht mang trn pha tnh th cc phn t nc c sp xp s gii

phng ra v t tp li thnh 1 khi nc.
ng dng ca phng php:
-

bc u tin cho qu trnh tinh sch khi mi trng

sau khi kt ta mui
sau khi chy sc k ion

c 3 bc ny u lun s dng nng mui cao.

HIC c s dng kt hp vi cc phng php tinh sch khc.
V d: Qu trnh tinh sch IgG1 ca chut quy m ln:
Mi trng t bolcb sung (NH4)2SO4HICc clc gel
Cu 15: Cc yu t nh hng n kt qu sc k tng tc k nc. Bn cht
ca pha tnh.
Tr li:
Cc yu t nh hng:
Pha tnh: - cht mang(ph thuc loi cht mang)
-cht kt gn (ph thuc loi, t trng ca cht kt gn)
Pha ng: - mui (loi mui, nng mui)
-pH
-nhit
-cht b sung
a. nh hng ca mui
S dng mui: gn nng mui cao, cht quan tm c gi li trn pha tnh; ra
gii nng mui thp.
nh hng ca mui n kt ta protein:
Xem danh sch cc ion trong slide

Cc ion lm tng cu trc ca nctng mnh ca tng tc k nc

nh hng ca mui n sc cng b mt ca nc
Danh sch cc mui xem trong slide

Tng sc cng b mt ca nc tng kh nng gi li ca protein trong HIC

S gi li ph thuc vo i lc ca mui vi pha tnh v cc protein
b.nh hng ca nhit v pH
nhit : kh nng gi li ca Pr tng theo nhit , ra tri Pr gim theo nhit
pH: s gi li ph thuc vo pI ca Pr, qu trnh ra gii c th thc hin bng
cch tng pH.
c. Cht b sung:
- tng tnh tan ca protein
- Tng qu trnh ra tri ca protein
- Ra ct
Water- miscible alcohol(ethanol, ethylenglycol)
Aliphatic amines (butylamine)
Detergents (triton X-100, Tween 20)
Chaotropic salts
Bn cht ca pha tnh:
Cc phi t k nc c c nh trn cht rn mang
Mc thay th c th nh hng n s phn tch
Cc loi vt liu c s dng:
Carbohydrates
-

Cross-link agarose (sepharose)

Dextran (sephadex)
Cellulose

Silica
Synthetic copolymer (polyme tng hp)
Cc cht gn:
Linear alkanes
Methyl<ethyl<propyl<butyl<pentyl<hexyl<octyl
Tnh k nc gia tng ca chui lm tng lc tng tc nhng cng c th lm gim tnh
chn lc hp ph
Aryl(aromatic) group: nhm cht thm
V d: phenyl
Kt hp tng tc k nc v tng tc ca cht thm (-)
Intermediate hydrophobic ligands: cac cht gn k nc trung gian
V d: polyethers
PEG(polyethylene glycol) < PPG(polypropylene glycol) < PTMG(polytetramethylene
glycol)
iu kin ra gii m du hn
Cu 16: Phn bit phng php sc k tng tc k nc v sc k o pha
Tr li:
HIC

RPC(reverse phase chrom)

Cht mang thay th t

Cht mang thay th nhiu

t cu t k nc

Nhiu cu t k nc

Lin kt yu

Lin kt mnh

Ra gii bng nc/ dung dch m long

Ra gii bng dung mi khng phn cc

Khng lm bin tnh protein

Lm bin tnh protein

Sc k hp ph

Sc k phn loi

Sc k di p sut thp

Sc k lng cao p (HPLC)

Cu 17: Mc ch to tinh th protein

- xc nh cu trc protein
- hiu c s lin kt ca protein vi cc cu t/ c cht/ thuc
- hiu c c ch xc tc enzyme ca protein
- xc nh v tr ng ca nguyn t trong phn t protein.
- Phng php: Tinh th hc tia X( 80% cu trc phn t protein c xc nh
bng tia X)
- Khng gii hn v kch thc: tt c virus v riboxom u c phn tch
- Tinh th dng khuch i tn hiu (1 tinh th cha 109 1013 phn t)
- S dng cho kt tinh phi hp vi cc cu t
Cu 18: Tnh cht ca tinh th protein
Khc vi phn t mui(sp xp cht), cc phn t protein c c tnh:
- Ln hn (>1000 nguyn t so vi 2- 10 nguyn t)
- Hnh dng khng u
- C b mt d thng. V vy cc tinh th protein c din tch tip xc gia cc
phn t ring bit t hn.
- Mm hn v d v hn
- Mt loi protein c th pht trin thnh cc tinh th c hnh dng khc nhau, s
dng cc vng tip xc gia cc phn t khc nhau.
- Cn c iu kin pH, nng mui, nhit gi trng thi ban u.
- Cha khong 50% nc hnh thnh cc knh
- Kh to tinh th hn mui nhiu.
Cu 19:
Cc giai on qu trnh tinh th ha protein
- Thu nhn lng ln protein tinh khit
- Chn lng dung dch m protein m protein c th tan tt v bn vng.
- Lm bo ha dung dch protein c th xy ra qu trnh to nhn tinh th mt
cch t pht
- Cc tinh th ln dn.
Cc yu t nh hng
- tinh khit ca protein
- Nng protein
- iu kin ban u (chun b dung dch protein)

Tc nhn gy kt ta
Nhit
pH
Cc cht b sung (cht ty ra, cht kh, c cht, ng nhn t)

C/ Cc phng php sy
1. Phn bit vt liu sinh hc v sn phm sinh hc.
2. Nhng bin i ca vi sinh vt trong qu trnh sy?
3. Phng php sy chn khng? ng dng?
4. phng php sy ng kh. ng dng?
5. phng php sy tng si. ng dng?
6. Phng php sy phun. ng dng?
7. Hy cho mt v d c th v vic sy tinh sch mt sn phm /vt liu sinh hc
(phn tch c s la chn phng php, iu kin thc hin qu trnh sy)

Cu 1:
- Vt liu sinh hc (biomaterial): l vt cht lin quan n c th sng hay chc
nng sng v hot ng sng.
V d: colagen, dextran, cellulose, chitosan, PLA, PGA
- Sn phm sinh hc (bioproduct): c nh ngha l 1 vt liu sinh hc hay l sn
phm ca qu trnh chuyn ha khi s dng vi sinh vt, cc thnh phn hay cc
cht c hot tnh sinh hc.
V d: etanol, enzyme
Cu 2:
Nhng bin i ca vi sinh vt trong qu trnh sy
- Bin i v tnh cht

Ha sinh

Enzyme

Ha hc

Vt l

S teo t bo (vi
khun, nm
men,
nm
mc)

Mt hot tnh
enzyme
v
vitamin

Gim gi tr dinh
dng
protein,
hydratcacbon,
cht bo, cht
khng sinh,
acid amin v
mt
hng
thm

Gim cc tnh
cht hu ch
(tnh tan, hi
nc, b co
li, gim khi
lng

Sy l qu trnh a vi sinh vt v dng tim sinh, l trng thi tm thi nhng

thun nghch v chc nng c to ra do tc ng ca cc yu t bn ngoi.
Trong trng thi ny, tc ca cc qu trnh din ra chm v n li tr li hot
ng bnh thng khi c iu kin tr li.

Cu 3:
Sy chn khng
- Sy chn khng ph hp cho cc vt liu d b bin i nhit nhng thng mt
thi gian sy ko di 20-100h. C th s dng sy chn khng lin tc v c th
t m 1% 40oC sau 10p.
- Nhit si 100oC 0,1Mpa, nhng 1kPa, si l 7 oC. Ngoi ra sy chn
khng gim c oxy ha.
- iu kin chn khng, nhit ha hi ca nc s rt thp, lm tng cng
qu trnh thot m trong vt, do vy c th tin hnh sy thp hn nhit mi
trng. V th, sn phm sy chn khng khng b tc ng gy bin tnh ca nhit
cao v lun gi c gn nh y cc tnh cht c trng ban u. Do ,
sn phm sy kh bng phng php ny gi c lu di v t b tc ng bi
iu kin bn ngoi.
ng dng:
- Sy vi khun lactic bng sy chn khng m bo sng st 52,8- 77,5
- Kt tinh bt ngt nhit thp
- Sy dc liu, phn hoa
- Sy g cng, g qu
Cu 4:
Sy ng kh

Sy ng kh l qu trnh lm lnh ng mu nhit -20 oC n -160oC, sau

lm bc hi nc bn trong v bn ngoi t bo di iu kin chn khng.
chn khng nh hn hoc bng 0,05mTorr v cc mu ng kh thng hoa nhanh.
- Sau 4-6h hn 95% lng nc t bo b loi b. Kh nng sng st ca t bo cao
- C th s dng cht bo v nh glycerol (to cho v t bo bn vng hn)
- u iem: Say nhiet o thap nen gio c cac tnh chat ti song
cua san pham. Neu dung e say thc pham seo gio c chat lng
va hng v cua san pham, khong b mat vitamin. Tieu hao nang lng
e bay hi am thap.
- Nhc iem: Gia thanh thiet b cao, van hanh phc tap, ngi van
hanh can co trnh o kyo thuat cao; tieu hao ien nang ln.
ng dng:
- Trong sn xut vi sinh, sy thng hoa c ng dng cho cc vi sinh vt, nm
men, vitamin, khng sinh, cc enzim khng bn nhit cao.
+ p dng rng ri trong sn xut dc phm ng kh cc cht khng sinh nh
peniciline, streptomycine v c bit l enzyme tinh khit
+S dng gi ging vi sinh vt.
- Say cac san pham myo pham, dc lieu, thc pham quy hiem.
- S dng trong cng ngh thc phm nh: sy rau qu (thanh long xut khu), tht
(tht heo ng kh s dng trong qun i)..
-

Cu 5:
Sy phun
- Thit b sy phun dng sy cc dng dung dch v huyn ph trong trng thi
phn tn nhm tch m ra khi vt liu gip tng bn v bo qun sn phm
c lu hn. Sn phm ca qu trnh sy phun l dng bt mn
- Dung dch cn sy kh lm m c bng cch bc hi chn khng di p
sut nht nh, a vo phn trn ca thng hnh tr y cn v c tuy-e thch hp
to tia lin tc di dng sng c ng knh 20-100m. ngang mc ca
tuy-e , mt lung kh nng(nhit khng vt qu nhit lm n nh sn
phm em sy), c phng vo v trn ln vi ht sng.
- Ty thuc vo nng sut tch m v sn phm c th ta c cc thng s k thut
thch hp. Tuy nhin, c cc thng s chung nh sau:
+ in th: 380V- 50Hz- 3 pha
+ Nhit sy: iu chnh t ng trong khong 30- 300oC
+ Ch lm vic: lin tc
+ iu khin: bo v chng qu nhit
- u im:
+ Qu trnh sy nhanh, c th iu khin c t trng sn phm.
+ Bt sau sy c ha tan cao (90-100%), m thp (3-4%)
+ Vn hnh lin tc v c th t ng ha hon ton.

+ Vn hnh v bo dng n gin, chi ph nhn cng thp

+ Thit k a dng cho tng loi sn phm, tng loi qui m nh my
+ p dng c cho cc sn phm bn nhit v khng bn nhit, nguyn liu dng
dung dch, gel, huyn ph
+ Cht lng bt c m bo trong sut qu trnh sy.
+ Vt liu hu nh khng tip xc vi b mt kim loi ca thit b
- Nhc im
+ Chi ph u t cao.
+ Yu cu m ban u cao m bo nguyn liu c th bm n thit b to git
lng.
+ Chi ph nng lng cao ( tch m)
+ Tht thot cc cht d b bay hi cao hn.
ng dng:
- S dng trong cng ngh thc phm nh: sn xut sa bt, bt chanh dy, lng
trng trng sy, caf, tr ha tan, bt rau qu, bt sa da.
- Sy phun trong cng nghip vi sinh c s dng sy kh cc cht c ca dung
dch canh trng cc cht khng sinh ng vt, cc axit amin, cc enzym, cc cht
trch ly nm thu nhn c trn cc mi trng dinh dng rn, cc dung dch
cht lng thu nhn c khi lm lng enzym bng cc dung mi v c hay bng
cc mui trung ho, cng nh cc phn c cht lng canh trng
- Sy phun cc ch phm probiotic c s dng cc cht mang-tc nhn bo v:
protein, polysaccarit, gum, maltodextrin, gelatin, tinh bt, lactoza, trehalose, skim
milk.
Cu 6:
Sy tng si
- L mt trong nhng k thut thng thng dng sy cc ht rn. K thut ny
da trn c s dng kh nng chuyn ng vi tc cao m to ra tng si vi
c tnh chuyn khi v truyn nhit.
- B phn chnh ca h thng sy tng si l mt bung sy, pha di bung sy l
b phn gia nhit. Khng kh c p sut ln v nhit thch hp c thi t b
phn gia nhit v lm cho ht dao ng nh l si.
- Vt liu sy trng thi si nhn nhit v nh m cho tc nhn tr nn nh hn,
theo tc nhn i ln lp trn v c ly ra ngoi mt cao thch hp m bo
ng kh yu cu.
- Dung e say cac vat lieu dang hat, o am trung bnh
- Cc thng s k thut thit b sy tng si:
+ Chiu cao vt liu tng si: 0,1-0,4m
+ Tc dng kh: 0,3-3,5m/s (ty thuc vo c tnh ca ht)
+ Chnh lch p sut: 300-1800 mmHg
- u im: Thiet b say tang soi co u iem la thch hp vi vat lieu

dang hat, o kho san pham rat ong eu, cng o say maonh liet do
s tiep xuc triet e gioa vat lieu va tac nhan say, cho phep say
nhiet o cao hn nhiet o cho phep v thi gian tiep xuc ngan. Hieu
suat s dung thiet b cao, gia thanh au t thap.
- Nhc im:
+ Chnh lch p sut tng i cao nn gy ra kh thi p lc cao.
+ Hnh dng ht b gii hn vi hnh cu hay hnh khi.
+ Kch thc ht b gii hn, vt liu phn tn gy ra hin tng cc vt liu tinh v
th khng to tng si tt.
+ Chiu cao ca tng si b gii hn bi chiu cao ti thiu m bo to to tng
si ng nht v chiu cao ti a cho php gim p sut.
+ Chu c tng si kt t s gy ra s khng n nh ca cc ht v c tnh chuyn
i gim.
- ng dng:
+ Sy qung, khong sn
+ Sy men bnh m
+ Sy thuc (sy bt, cm trong nguyn liu dc phm, thc phm, ha cht)