Chng 4

Thit k h ln men
I. H ln men thng khuy
Ni phn ng sinh hc (bioreactor) hay cn gi l h ln men (fermenter) l loi thit b m trong n s bin i ha sinh c tin hnh bi cc t bo sng hoc cc thnh phn t bo in vivo (enzyme). Trong chng ny, ni phn ng sinh hc nui cy cc t bo sng c gi l h ln men phn bit cc ni phn ng sinh hc dng cho cc enzyme. Trong phng th nghim, cc t bo thng c nui cy trong cc bnh tam gic trn my lc. Lc nh bnh tam gic rt hiu qu to ra dch huyn ph t bo, tng cng s oxy ha thng qua b mt cht lng v tr gip s chuyn khi (mass transfer) ca cc cht dinh dng m khng gy nguy him cho cu trc t bo.

bt

vch ngn un nng lm lnh turbin dt

khng kh v trng

Hnh 4. 1. S h ln men dng cho sn xut penicillin.

i vi hot ng sn xut quy m ln, th h thng ln men thng khuy (stirred-tank fermenter, STF) c s dng rng ri nht thit k cho qu trnh ln men cng nghip. N c th c dng cho c hai trng hp ln men hiu kh (aerobic) v ym kh (anaerobic) trong mt phm vi rng cc loi t bo khc nhau bao gm vi sinh vt, ng vt v thc vt.

Cng ngh t bo

33

Hnh 4.1 gii thiu s h ln men dng trong sn xut penicillin. Cng pha trn (mixing intensity) c th rt khc nhau bng cch chn loi cnh khuy (impeller) thch hp v cc tc khuy khc nhau. Vic sc kh v khuy c hc trong h ln men rt tt cho nui cy dch huyn ph t bo, s oxy ha, s pha trn mi trng v truyn nhit. STF cng c th c dng cho cc mi trng c nht cao. N l mt trong nhng h ln men quy m ln u tin c pht trin trong cng nghip dc. c im v tim nng ca STF c nghin cu rng ri. Do h ln men thng khuy thng c lm bng thp khng r v hot ng trong iu kin n ha nn tui th ca thit b rt lu. Nhc im ca h ln men thng khuy bt ngun t u im ca n. B phn (cnh) khuy rt hiu qu trong vic pha trn cc thnh phn ca h ln men, nhng li tiu th mt lng ln cng sut v c th gy nguy him cho nhng h thng t bo nui cy mn cm vi lc trt (shear force) nh t bo ng vt c v hoc t bo thc vt. Lc trt ca cht lng trong hn hp c to ra bi gradient tc ca cc thnh phn tc (hng tm v tip tuyn) ca cht lng khi ri khi vng cnh khuy. Khi cht lng ri khi vng trung tm, th tc ca n v tr trn v di cnh khuy (c khong cch bng chiu rng cnh khuy) s gim khong 85% v to ra mt vng trt cao. Khi t l chiu rng cnh khuy trn ng knh ca n tng th profile tc t c dng c trng ca parabol m tr nn t hn v n to ra lc trt t hn do gradient tc ln dn ln. V th, bng cch tng chiu rng cnh khuy, c th ng dng thnh cng STF trong nui cy t bo ng vt hoc t bo thc vt. Nhiu h ln men quy m phng th nghim c lm bng thy tinh c np bng thp khng r. Cc thng ln men ln hn c lm bng thp khng r. T l chiu cao trn ng knh ca thng ln men (vessel) hoc l 2/1 hoc l 3/1 v thng c khuy bng hai hoc ba turbine khuy (cnh khuy). Trc cnh khuy c gn trn np hoc t y ca thng bng gi . T l ng knh cnh khuy (DI) trn ng knh ca thng (DT) thng l t 0,3-0,4. Trong trng hp h ln men c hai cnh khuy, th khong cch gia cnh khuy th nht vi y ca vessel v khong cch gia hai cnh khuy bng 1,5 ng knh cnh khuy. Khong cch ny gim xung cn 1,0 so vi ng knh cnh khuy trong trng hp h ln men c ba cnh khuy. Bn vch ngn (baffles) cch u nhau thng

Cng ngh t bo

34

c thit k ngn cn s hnh thnh dng xoy lm gim hiu sut pha trn. Chiu rng ca vch ngn thng bng 1/10 ng knh ca thng (tank). trng hp h ln men hiu kh (aerobic fermenter), th mt b phun l n (single orifice sparger) hoc mt b phun vng c s dng sc kh cho h ln men. B phn phun c t v tr gia cnh khuy cui cng v y ca vessel. pH trong h ln men c th c duy tr bng cch dng dung dch m hoc b iu chnh pH (pH controller). Nhit c iu chnh bng h thng gia nhit v lm lnh t ng. 1. H ln men dng nt (plug-flow fermenter, PFF) hoc m (batch fermenter) Mt h ln men khuy l tng phi c kh nng pha trn tt sao cho cc thnh phn ng nht trong mt kt cu mi thi im. Mt h ln men l tng khc l h ln men dng nt, mt dng tng ng ca h ln men m. Trong h ln men dng ng (tubular-flow fermenter), cht dinh dng (c cht) v t bo i vo mt u ca ng hnh tr v t bo s sinh trng trong khi chng i qua ng ny. Do ng di v thiu b phn khuy nn ngn cn s pha trn hon ton ca cht lng, v th tnh cht ca dng chy thay i trong hai chiu tip tuyn v hng tm. Tuy nhin, s bin thin trong chiu hng tm nh hn chiu tip tuyn. Mt h ln men dng ng m khng c nhng bin thin hng tm th c gi l h ln men dng nt (PFF). Thc t, h ln men PFF rt kh xy dng. Cho d h ln men PFF trng thi n nh (steady state) c hot ng trong mt kiu lin tc, th nng t bo ca h ln men m l tng sau thi gian t s ging nh nng t bo ca h ln men PFF trng thi n nh v tr chiu dc ni m thi gian lu (residence time) bng t (Hnh 4.2). V th, s phn tch sau y ng dng cho c hai, h ln men m l tng v PFF trng thi n nh. Nu mi trng lng c tip mu bng nui cy kt ht (seed culture), th t bo s bt u sinh trng theo hm m sau pha lag. Trong h ln men m, s thay i nng t bo bng tc sinh trng t bo:

dC X = r X = C X dt

(4.1)

Cng ngh t bo

35

F
V, CX , CS

CXo Cso to t CX Cs
(b)

CX f CS f

C X = CX o Cs = Cso
(a)

t = to

Hnh 4.2. S (a) h ln men thng khuy m v (b) h ln men dng nt.

thu c phng trnh hiu sut ca ln men m, chng ta cn ly tch phn phng trnh (4.1) s c:
CX

CX 0

t X dC X dC X = = dt = t t 0 rX C X t0 CX
0

(4.2)

Cn lu rng, phng trnh (4.2) ch c ng dng khi rX > 0. V th, t 0 (trong phng trnh 4.2) khng phi l thi gian ca nui cy ban u sau khi tip mu, m l thi gian t bo khi ng sinh trng, l giai on pha sinh trng bt u tng nhanh. Theo phng trnh (4.2), thi gian sinh trng tng m t t 0 chnh l din tch pha di ng cong 1/rX theo C X gia C X 0 v C X (Hnh 4.3). ng cong lin tc hnh 4.3 c tnh ton bng phng trnh Monod v vng c mu ti bng t t 0 . Thi gian sinh trng tng m t khi c c lng bng th ny v xc nh n th da vo ng cong t theo C X l n gin hn. Tuy nhin, biu din bng th s thun tin trong vic so snh tim nng ca cc cu hnh h ln men khc nhau (s c tho lun sau). Lc ny ch lu rng, ng cong c mu ti dng ch U l c trng ca cc phn ng xc tc t ng: S+XX+X
Cng ngh t bo 36

1 rX
1

CX
Hnh 4.3. th ca thi gian sinh trng tng m t t 0 (vng ti). ng cong lin tc biu din m hnh Monod vi max = 0,935/gi; K S = 0,71 g/L;

YX/S = 0,6; C X 0 = 1,6 g/L; v CS 0 = 10 g/L. Tc khi u ca phn ng xc tc t ng chm do nng ca X thp. Tc phn ng tng ln khi cc t bo sinh sn v sau s t n tc ti a. Khi lng c cht gim v cc sn phm c c tch ly, th tc phn ng gim xung gi tr thp hn. Nu ng hc Monod (Monod kinetics) biu din thch hp tc sinh trng trong sut pha hm m, th chng ta c th thay th phng trnh (3.11) chng 3 vo phng trnh (4.2) c c:
CX

CX 0

t ( K S + C S )dC X = dt max C S C X t0

(4.3)

Phng trnh (4.3) c th tnh c tch phn nu chng ta bit mi quan h gia CS v CX. Ngi ta quan st thy rng s lng sinh khi t bo c sn xut t l vi lng c cht gii hn c tiu th. Hiu sut sinh trng ( Y X/S ) c nh ngha nh sau:

YX/S =

C X CX0 C X = C S (C S C S0 )

(4.4)

Thay phng trnh (4.4) vo phng trnh (4.3), tch phn ca phng trnh tng hp ny s a ra mi quan h gia nng t bo v thi gian:
Cng ngh t bo 37

(t t 0 ) max

C CS K S YX / S K S YX / S = + 1 ln X + ln 0 C X + C S YX / S CX C X 0 + C S0 YX / S CS 0 0 0

(4.5)

2. H ln men thng khuy lin tc (continuous stirred-tank fermenterCSTF) l tng Qun th t bo c th tip tc giai on sinh trng hm m trong mt thi gian di bng cch duy tr h thng nui cy lin tc. Hnh 4.4 trnh by s h ln men thng khuy lin tc (CSTF). Bung sinh trng (thng ln men hay bnh nui) c kt ni vi bnh cha mi trng v trng. Khi qu trnh sinh trng bt u th mi trng sch c cung cp lin tc t bnh cha mi trng. H thng nui cy lin tc c th hot ng nh l mt chemostat (th n nh ha tnh) hoc turbidostat (th n nh c). Trong chemostat tc dng chy c ci t mt gi tr c bit v tc sinh trng ca nui cy s iu chnh tc dng chy ny. Ni chung, hot ng chemostat d dng hn turbidostat, do n c th c thc hin bng cch t my bm mt tc dng chy khng i, trong khi turbidostat i hi mt thit b cm quang (optical sensing device) v mt b iu chnh (controller). Tuy nhin, turbidostat c gii thiu khi h ln men lin tc cn c tin hnh cc tc pha long cao gn vi im ra tri (washout point), khi ta c th ngn cn s ra tri bng cch iu ha tc dng chy trong trng hp tht thot t bo thng qua dng chy ra ngoi vt qu s sinh trng t bo trong h ln men.

CXi Csi

V, CX , CS

CX F C s

Hnh 4.4. S h ln men thng khuy lin tc (CSTF).

Cng ngh t bo

38

Cn bng nguyn liu cho t bo trong CSTF (Hnh 4.4) c th c vit nh sau:

FC X i FC X + VrX = V

dC X dt

(4.6)

Trong : rX l tc sinh trng t bo trong h ln men v dC X / dt biu din s thay i nng t bo trong h ln men theo thi gian. i vi CSTF hot ng trng thi n nh, th s thay i nng t bo theo thi gian l bng khng (dCX / dt = 0) do cc t bo trong bnh nui ch sinh trng nhanh thay th nhng t bo b hao ht theo dng chy ra ngoi, v phng trnh (4.6) tr thnh:

m =

V C X C Xi = F rX

(4.7)

Phng trnh (4.7) cho thy thi gian lu cn thit (m) bng din tch hnh ch nht c chiu rng C X C X i v chiu cao 1 / rX trn ng cong

1 / rX theo CX.
Hnh 4.5 biu din ng cong 1 / rX theo CX. Din tch hnh ch nht c t m trong hnh bng thi gian lu trong CSTF khi dng chy vo l v trng. Minh ha thi gian lu bng th c th gip chng ta so snh hiu qu ca cc h ln men. H ln men c thi gian lu ngn hn ( t ti mt nng t bo nht nh) l hiu qu hn. Hot ng ti u ca h ln men da trn s minh ha th ny s c tho lun trong phn tip theo.
4

1 rX

3 2 1

CX

Hnh 4.5. Minh ha bng th c lng thi gian lu cho CSTF. ng biu din m hnh Monod vi max = 0,935/gi; K S = 0,71 g/L; Y X/S = 0,6;

CS i = 10 g/L; v C X i = 0.
Cng ngh t bo 39

Nu dng chy vo l v trng (C X i = 0), v t bo trong CSTF ang sinh trng theo hm m (rX = C X ) th phng trnh (4.7) s tr thnh: 1 1 D

m =

(4.8)

Trong : D c bit nh l tc pha long v c gi tr bng nghch o ca thi gian lu ( m ). V th, i vi CSTF trng thi n nh c cht dinh dng v trng, th tc sinh trng c trng bng tc pha long. Mt khc, tc sinh trng c trng ca t bo c th c iu chnh bng cch thay i tc dng chy mi trng. Nu tc sinh trng c th c biu din bng phng trnh Monod, th sau :

D==

max C S
K S + CS

(4.9)

T phng trnh (4.9), CS c th c tnh ton bng thi gian lu bit v cc thng s ng hc Monod nh sau:

CS =

KS

m max 1

(4.10)

Tuy nhin, cn ch rng phng trnh (4.10) ch c gi tr khi m max > 1 . Nu m max < 1 , tc sinh trng ca t bo s thp hn tc t bo tht thot theo dng chy ra ngoi. Do , tt c t bo trong h ln men s b ra tri, v phng trnh (4.10) s khng c gi tr. Nu hiu sut sinh trng (YX / S ) l hng s, th sau :

C X = YX / S (C Si CS )

(4.11)

Thay phng trnh (4.10) vo phng trnh (4.11) s cho hiu sut tng quan i vi CX nh sau:

KS C X = YX / S C S i 1 m max

(4.12)

Cng ngh t bo

40

Tng t:

KS CP = CPi + YP / S C S i 1 m max

(4.13)

Trong : CP l nng sn phm, CPi l nng sn phm a vo. Mt ln na, phng trnh (4.12) v (4.13) ch c gi tr khi m max > 1 . Trong phn ny, chng ta t cn bng nguyn liu cho nng t bo v thu c cc phng trnh khc nhau cho CSTF. Cc phng trnh tng t cng c th thu c bng cch t cc cn bng nguyn liu cho nng c cht v nng sn phm. 3. c lng cc thng s ng hc Monod ng thc tc sinh trng c trng v tc pha long ca CSTF trng thi n nh (phng trnh 4.9) tin li trong nghin cu nh hng ca cc thnh phn khc nhau ca mi trng ln tc sinh trng c trng. Bng cch o nng c cht trng thi n nh vi cc tc dng chy khc nhau, cc m hnh ng hc khc nhau c th c th nghim v gi tr ca cc thng s ng hc c th c c lng. Sp xp li phng trnh (4.9) c th thu c mi quan h tuyn tnh nh sau:

max

KS

1 1 + CS max

(4.14)

Trong : bng tc pha long (D) cho chemostat. Nu mt t bo nht nh tun theo ng hc Monod, th th 1 / theo 1 / CS s em li gi tr max v KS (bng cch c phn b chn v dc ca ng thng). th ny c u im cho thy mi quan h gia bin c lp (CS) v bin ph thuc . Tuy nhin, 1 / s tin ti nu nng c cht gim dn n trng lng vt qu mc o khi nng c cht thp v trng lng khng o khi cc nng c cht cao. Phng trnh (4.9) c th sp xp li a ra cc mi quan h tuyn tnh ng dng thay cho phng trnh (4.14) nhm c lng tt hn cc thng s trong nhng trng hp nht nh:
Cng ngh t bo 41

CS

KS

max

CS

max

(4.15)

= max K S

CS

(4.16)

Tuy nhin, gii hn ca php tnh gn ng ny ( xc nh cc thng s ng hc) gp kh khn khi s dng CSTF. i vi trng hp vn hnh theo tng m, chng ta thm ch c th dng bnh tam gic lc trn my lc vn hnh nhiu m vi cc iu kin khc nhau trong cng mt thi gian. Vn hnh theo tng m trong ni ln men c khuy cng khng kh khn lm, do khng c cc kt ni i vo v i ra (ngoi tr b phn cung cp khng kh) v thi gian vn hnh ngn, t c nguy c ca s nhim bn h ln men. vn hnh CSTF, chng ta cn c cc ngun cung cp dinh dng v tch tr sn phm c kt ni v trng vi h ln men. Tc ca cc dng chy vo v ra khi h ln men cn c kim sot mt cch chnh xc. Thnh thong, vic kim sot tc dng chy ra c th gp kh khn do s to bt v kt khi ca cc t bo. Do thi gian vn hnh t nht mt vi ngy hoc thm ch c tun t ti trng thi n nh (cng gy ra s bin i tc pha long), cho nn lun c ri ro cao i vi h ln men do b nhim bn. Thng xuyn gp kh khn trong vic t ti trng thi n nh bi t bin ca t bo v kh nng thch nghi vi mi trng mi ca chng. Hn na, do hu ht cc h ln men quy m ln c tin hnh trong kiu tng m, cho nn cc thng s ng hc c xc nh bi nghin cu chemostat phi d bo c s sinh trng trong kiu ln men ny. Tuy nhin, bng chng (kim tra v xc minh) m hnh ng hc v c lng cc thng s ng hc bng cch vn hnh chemostat l phng php ng tin cy nht do iu kin mi trng khng thay i ca n. Cc s liu ca vn hnh theo tng m c th c dng xc nh cc thng s ng hc, cho d n khng phi l phng thc c gii thiu cao. Tc sinh trng c trng trong sut qu trnh vn hnh theo tng m c th c c lng bng cch o dc ca ng cong nng t bo theo thi gian cc im khc nhau. Nng c cht cn thit c o cng cc im ni m dc c c. Sau cc th theo cc phng trnh (4.14), (4.15) v (4.16) c th c xy dng xc nh
Cng ngh t bo 42

cc thng s ng hc. Tuy nhin, gi tr ca cc thng s thu c trong phng php ny cn thit c kho st cn thn xem chng c trong phm vi hp l cho cc t bo c kim tra hay khng. 4. Hiu sut ca CSTF Thng thng, hiu sut ca h ln men c hiu nh l s lng sn phm c sn xut trn mt n v thi gian v th tch. Nu dng chy vo l v trng (C X i = 0) th hiu sut sinh khi t bo bng C X / m , chnh l dc ca ng thng OAB ca ng cong CX theo m (Hnh 4.6).

10 8 6

CX
4 2

A O
0

Hnh 4.6. S thay i nng t bo v c cht nh l mt hm ca thi gian lu. Hiu sut bng dc ca ng thng OAB . ng cong c v bng m hnh Monod vi max = 0,935/gi; KS = 0,71 g/L; YX/S = 0,6; v C S i = 10 g/L.

Hiu sut im A bng hiu sut im B. im A nng t bo ca dng chy ra thp nhng thi gian lu li ngn, v th mi trng c th chy qua d dng hn. Ngc li, im B nng t bo ca dng chy ra cao nhng thi gian lu li di v th ch c mt lng nh ca mi trng chy qua. im A l vng khng n nh v rt gn vi im ra tri D, v v ch cn mt s dao ng nh trong thi gian lu cng c th em li mt s thay i ln trong nng t bo. Khi dc ca ng thng
Cng ngh t bo 43

tng ln th hiu sut s tng v di ca AB gim. dc ca ng thng s t gi tr cc i khi n l ng tip tuyn ca ng cong CX. V th, gi tr hiu sut cc i bng dc ca ng OC . Hiu sut cc i s t c im D. iu kin hot ng t hiu sut cc i CSTF c th c lng theo th bng cch dng ng cong 1 / rX theo CX. Hiu sut cc i c th thu c khi thi gian lu l ti thiu. V thi gian lu bng din tch ca hnh ch nht vi chiu rng CX v chiu cao 1 / rX trn ng cong 1 / rX theo CX, cho nn n s t ti thiu khi 1 / rX l ti thiu (Hnh 4.7).
4

1 rX

3 2 1

CX

Hnh 4.7. Minh ha bng th CSTF vi hiu sut cc i. ng lin tc biu din cho m hnh Monod vi max = 0,935/gi; KS = 0,71 g/L; YX/S = 0,6; C Si = 10 g/L; v C X i = 0 .

iu cn lu l iu chnh cc phng trnh cho nng t bo v thi gian lu sao cho hiu sut t bo t cc i. Hiu sut t bo cho CSTF trng thi n nh vi cht dinh dng v trng l:

CX

= rX =

max C S C X
K S + CS

(4.17)

Hiu sut t cc i khi drX / dC X = 0, sau khi thay th CS = CS i C X / YX / S vo phng trnh (4.17), ly tch phn theo CX v t phng trnh tng hp bng 0, chng ta thu c nng t bo ti u (C X , opt ) cho hiu sut cc i nh sau:
Cng ngh t bo 44

C X , opt = YX / S CS i

+1

(4.18)

Trong :

K S + C Si KS

(4.19)

V: C S = C Si

CX nn nng c cht ti u (CS, opt) s l: YX / S

C S ,opt = C Si
(4.20)

+1

Thay phng trnh (4.20) vo phng trnh (4.17) thu c mt thi gian lu ti u (m,opt) nh sau:

m ,opt =

max ( 1)

(4.21)

5. So snh nui cy ca h ln men m v h ln men thng khuy lin tc Nh cp, thi gian lu cn thit nui cy m hoc PFF trng thi n nh t ti mt nng t bo nht nh l:

b = t0 +

CX

CX0

dC X rX

(4.22)

Trong : t0 l thi gian cn thit t ti pha sinh trng theo hm m. Din tch bn di ca ng cong 1 / rX theo CX, gia C X i v CX l bng b t1 nh c trnh by hnh 4.3. Mt khc, thi gian lu CSTF c biu din bi phng trnh (4.17) bng din tch hnh ch nht vi chiu rng C X C X i , v chiu cao

1 / rX .

Cng ngh t bo

45

V ng cong 1 / rX theo CX c dng hnh ch U nn chng ta c th c mt vi nhn xt cho h ln men n nh sau: - Hu ht cc h ln men sn xut l mt CSTF hot ng vi nng t bo m gi tr ca 1 / rX l ti thiu (Hnh 4.8 a) do n i hi thi gian lu ngn nht. - Nu nng cui cng ca t bo c hng ti trong pha tnh, th h ln men m l chn la tt hn CSTF, v thi gian lu cn thit cho nui cy m (Hnh 4.8 b) l ngn hn ca CSTF.

1 rX

1 rX

CX

CX

Hnh 4.8. Minh ha bng th thi gian lu c yu cu (vng ti) cho: (a) CSTF v (b) h ln men m.

II. Thu hi t bo
i vi hot ng lin tc ca PFF v CSTF, cc t bo tht thot cng vi dng chy ra (outlet) hn ch hiu sut ca h ln men. V th, hiu sut c th c ci thin bng cch thu hi (recycling) t bo t dng chy ra a tr li h ln men. 1. Thu hi t bo PFF PFF i hi s hin din ban u ca t bo trong dng chy vo (inlet) nh l mt h ln men m i hi a mu vo ban u. Phng thc kinh t nht cung cp t bo trong dng chy vo l thu hi mt phn ca

Cng ngh t bo

46

dng chy ra a tr li dng chy vo vi (hoc khng c) thit b tch ri t bo. Hnh 4.9 m t s thu hi t bo PFF. Khng ging nh CSTF, PFF khng i hi thit b tch ri t bo thu hi, v s hin din ca n khng lm tng ng k hiu sut ca h ln men. Phng trnh hiu sut ca PFF vi ng hc Monod c th c vit nh sau:

p V = = (1 + R ) F 1 + R

CX f

' CX

dC X = rX

CX f

' CX

( K S + C S )dC X max C S C X

(4.23)

Trong : p l thi gian lu da trn tc dng chy ca ton b h thng. Thi gian lu thc t trong h ln men ln hn p do tc dng chy tng ln nh thu hi t bo. (1 + R)F CX Cs RF CXR , Csf

CXi F Csi

CXf Csf CXL = 0 Csf

Hnh 4.9. S thu hi t bo PFF.

Nu hiu sut sinh trng l khng i th:

' CS = CS

Y X/S

' (C X C X )

(4.24)

Thay phng trnh (4.24) vo trong phng trnh (4.23) cho CS v ly tch phn ta s c kt qu sau:

p max

' CX f K S Y X/S K S YX / S CS 1 ln ln + = + ' ' C' C' + C' Y 1+ R CX YX / S C S f + CS S X/S X X

(4.25)

Cng ngh t bo

47

' ' Trong : C X v C S c th c c lng t s cn bng t bo v

c cht im phi trn ca dng chy vo v dng chy thu hi nh sau:

' CX =

C X i + RC X R 1+ R C Si + RC S R 1+ R

(4.26)

' CS =

(4.27)

Nng t bo ca dng chy ra, c th c c lng t ton b s cn bng t bo nh sau: CX f = 1

[C

Xi

+ Y X/S (C Si C S f )

(4.28)

Nng t bo ca dng chy thu hi c th c c lng t s cn bng t bo trn b lc nh sau:

CXR =

1+ R CX f R

(4.29)

Trong : l t l x (bleeding) c nh ngha nh sau:

B F

(4.30)

Hnh 4.10 trnh by hiu qu ca tc thu hi (R) trn thi gian lu ca h thng PFF c thu hi. Lu rng thi gian lu c tnh ton da trn tc dng chy vo, l thi gian lu thc s ca h ln men. Thi gian lu thc t trn h thng PFF l khng quan trng bi v n s gim xung khi tng tc thu hi. Khi = 1, tc x s bng tc dng chy, v tc dng chy ca phn c lc L l bng 0, v th dng chy thu hi khng c lc. Thi gian lu s l v hn nu R bng 0 v gim r rt khi R tng ln. Trong trng hp ny t l thu hi ti u c th trong khong 0,2. Mt ng cong khc trong hnh 4.10 l cho = B / F = 1,8. Thi gian lu cn thit c th gim bng cch tp trung dng chy thu hi t 2540% khi R trong khong 0,2-1,0. Khi R 1,2, th mt on ca ng

Cng ngh t bo

48

cong c biu din bng du chm, bi v kh c th gim t l thu hi xung di 0,2 khi = 0,8.
8

p =

V F

=1
2

0 0,0

= 0,8
0,2 0,4 0,6 0,8 1,0

R
Hnh 4.10. nh hng ca tc thu hi (R) v t l x ( = B / F ) ln thi gian lu (p = V/F gi) ca h thng PFF c thu hi. ng cong c v bng m hnh Monod vi max = 0,935/gi; KS = 0,71 g/L; YX/S = 0,6; C S i = 10 g/L; C S f = 1,3 g/L; v C X i = 0.

Vic phn tch trong phn ny v cc phn sau cng c th c ng dng trong cc b lng t bo nh l mt b phn phn tch t bo. Dng chy ra ca b lng t bo s bng F = B+L v nng ca n s l ( B / F ) C X f = C X f . 2. Thu hi t bo CSTF Hiu sut t bo trong CSTF tng ln cng vi vic tng tc pha long v t n gi tr cc i. Nu tc pha long tng ln qu im cc i, th hiu sut ca h ln men s gim t ngt v t bo s bt u b pha long do tc sinh sn t bo km hn s hao ht t bo dng chy ra. Mt phng thc ci thin hiu sut h ln men l thu hi t bo bng cch tch ri t bo khi dng chy sn phm bng h lc dng chy ngang (cross-flow filter unit) (Hnh 4.11). Nng cao ca t bo (c duy tr bng cch thu hi t bo) s lm tng hiu sut t bo khi tc sinh trng t l tng ng vi nng t
Cng ngh t bo 49

bo. Tuy nhin, phi c gii hn trong vic tng hiu sut t bo vi vic tng nng t bo bi v trong mi trng c nng t bo cao, th tc chuyn khi cht dinh dng s b gim do vic dn vo mt ni qu ng v gy kt khi ca t bo. Vic duy tr nng qu cao ca t bo cng khng c li bi v b phn lc s thng xuyn b hng hn trng hp nng t bo cao.

CXi Csi

L V B
Hnh 4.11. S thu hi t bo CSTF.

CXL = 0 Csf CX f Csf

Nu tt c t bo c thu hi tr li trong h ln men, th nng t bo s tng lin tc theo thi gian v trng thi n nh s khng bao gi t c. V th, hot ng ca CSTF c s thu hi trng thi n nh, chng ta cn c mt dng x (Hnh 4.11). Phng trnh cn bng nguyn liu cho t bo trong h ln men c b phn thu hi t bo c dng nh sau:

FC X i BC X + VC X = V

dC X dt

(4.31)

Cn lu rng, tc dng chy thc t i vo v i ra khi b phn lc khng quyt nh hon ton n s cn bng tt c nguyn liu. i vi CSTF trng thi n nh c s thu hi t bo v cht dinh dng v trng, th:

D =

= m

(4.32)

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50

Lc ny D thay cho D v bng tc sinh trng c trng. Khi D = 1 t bo khng c thu hi, v th = . Nu tc sinh trng c th biu din bng ng hc Monod, th thay th phng trnh (3.11) chng 3 vo phng trnh (4.32) ta c:

CS =

K S m max

(4.33)

CS ch c ngha khi m max > . Nng t bo trong h ln men c

th c tnh ton t gi tr ca CS nh sau:

CX =

YX / S

(C Si C S )

(4.34)

Hnh 4.12 cho thy nh hng ca t l x ln hiu sut t bo i vi m hnh Monod. Khi b gim xung t 1 (khng thu hi) ti 0,5 th hiu sut t bo c tng ln gp i.
10 8

= 0,05
Khng thu hi

DC X 6
4 2 0 0,0

Hnh 4.12. nh hng ca t l x ln hiu sut t bo (DCX).

0,5

1,0

1,5

2,0

III. Cc h ln men khc

Nhiu h ln men khc c xut v th nghim. Cc h ln men ny c thit k ci thin hoc nhc im ca h ln men thng khuy (tiu th cng sut ln) hoc cc yu cu c bit ca mt qu trnh ln men nht nh nh: sc kh tt hn, chuyn nhit hiu qu, tch hoc gi li t bo, bt ng t bo, gim bt thit b v gi thnh ca sn phm, v thng khng c thit k cho quy m ln.

Cng ngh t bo

51

Cc h ln men thng c phn loi da trn c s cc kiu bnh nui ca chng nh l thng, ct, hoc cc h ln men vng (loop). C hai h ln men thng v ct c xy dng trn c s bnh nui hnh tr. C th phn loi da theo t l chiu cao (H) trn ng knh (D) nh sau: H/D < 3 cho h ln men thng. H/D > 3 cho h ln men ct. H ln men vng l h ln men thng hoc ct c vng lu thng cht lng, (c th l ng thng gi (draft) gia hoc l mt ci vng gn bn ngoi h ln men). C th phn loi cc h ln men theo mt cch khc da trn c s cc thnh phn ca h ln men c phi hp nh th no: bi kh nn, bi b phn chuyn ng c hc bn trong, hoc bi bm cht lng bn ngoi. Cc h ln men tiu biu trong mi loi c trnh by bng 4.1, v cc u im v nhc im ca ba loi h ln men c bn c trnh by bng 4.2.
Bng 4.1. Phn loi cc h ln men. Ngun trn s cp Kh nn Ct bong bng Ct hnh nn Vng Cc b phn chuyn ng bn trong Thng khuy Nhiu giai on (hoc t) Bm bn ngoi Khay sng (ry) m nhi Vng tia

Loi bnh nui Thng Ct

p lc khng kh Vng chn vt y ln theo chu k

1. H ln men ct (column fermenter) H ln men n gin nht l h ln men ct bong bng (cn gi l h ln men thp-tower fermenter), thng bao gm mt bnh tr di, c b phn phun kh di y (Hnh 4.13 a-c). Cc thnh phn ca h ln men c trn bng cch tng s lng bong bng ln, cng l yu t c th cung cp oxygen cn thit cho t bo. Khi cc t bo lng xung, nng

Cng ngh t bo

52

cao ca t bo c th c duy tr phn thp hn ca ct m khng c bt k mt thit b no tch ri chng. Tuy nhin, h ln men ct bong bng thng b hn ch trng hp ln men hiu kh v vic tng cc bong bng khng th cung cp mt s pha trn y cho s sinh trng ti u. Ch c phn thp hn ca ct c th duy tr nng t bo cao dn n s ln men ban u nhanh c tip theo bi s ln men chm hn do cc c cht mong mun b gim i. Khi nng t bo tng ln trong h ln men, cn c lu tc khng kh cao duy tr dch huyn ph t bo v s pha trn. Tuy nhin, lu tc khng kh tng ln c th gy ra s to bt nhiu v vic duy tr cc bong bng kh trong ct dn n lm gim hiu sut ca h ln men. Khi bong bng tng ln nhiu trong ct chng c th kt thnh mt khi nhanh chng lm gim tc chuyn oxygen. V th, cc h ln men ct c th khng thay i c v hn ch mt phm vi kh hp cc iu kin hot ng.
Bng 4.2. u im v nhc im ca cu hnh ba h ln men c bn. Loi Thng khuy u im 1. Linh hot v d thch ng 2. Phm vi mt pha trn rng Nhc im 1. Tiu th cng sut ln

2. Gy tn hi cho cc t 3. C th s dng mi trng c bo mn cm vi lc trt 3. Gi thnh thit b cao nht cao 1. Khng c cc b phn chuyn ng 1. Pha trn km 2. To bt d tha 3. Gii hn i vi h thng c nht thp 1. Pha trn km 2. To bt d tha 3. Gii hn i vi h thng c nht thp

Ct bong bng

2. n gin 3. Gi thnh thit b thp 4. Nng t bo cao 1. Khng c cc b phn chuyn ng 2. n gin 3. Hiu sut ht kh cao 4. Chuyn nhit tt

Lc y khng kh

khc phc nhc im ca h ln men ct, mt vi kiu thit k khc c xut. H ln men ct hnh chp ngc (Hnh 4.13 b) c th

Cng ngh t bo

53

duy tr lu tc khng kh cao trn mt n v din tch phn thp hn ca h ln men m c nng t bo cao. Mt vi khay sng lc c th c ci t trong ct (Hnh 4.13 c) tng hiu qu tip xc kh-cht lng v ph v s kt khi ca bong bng kh. tng cng s pha trn m khng c cc phn chuyn ng bn trong, dch ln men (mi trng) c th c bm ra ngoi v quay vng (tun hon) bng cch dng mt bm cht lng bn ngoi (Hnh 4.13 d v e).

Khng kh (a) (b) (c) (d) (e)

Hnh 4.13. Cc h ln men ct: (a) ct bong bng (bubble column), (b) ct hnh nn (tapered column), (c) ct bong bng c khay sng lc (sieve-tray bubble column), (d) ct bong bng c khay sng lc vi bm bn ngoi, (e) ct nhi (packed-bed) vi bm bn ngoi.

2. H ln men vng (loop fermenter) H ln men vng l h ln men thng (tank fermenter) hoc ct (column fermenter) c vng lu thng cht lng, n c th l mt ng thng gi gia hoc l mt ci vng bn ngoi. Ty thuc vo s lu thng cht lng c to ra nh th no, m h ln men c phn loi thnh ba kiu: lc y khng kh (air-lift), vng khuy (stirred loop) v vi phun (jet loop) (Hnh 4.14). S lu thng cht lng ca h ln men dng lc y nh vo vic phun khng kh to ra s khc nhau v mt gia phn giu bong bng ca cht lng trong tm ng (riser) v phn c rt ht bong bng nng hn ca cht lng trong y thng (downcomer) (Hnh 4.14 a). S pha trn v lu thng cht lng trong thng ln men c th c tng cng bng cch gn thm mt b phn bm bn ngoi (Hnh 4.14 b). Tuy nhin, vic b sung
Cng ngh t bo 54

bm lm gim u im ca h ln men nh lc y khng kh l hiu sut nng lng thp v n gin. H ln men p lc chu k ICI (Imperial Chemical Industries Ltd., England) l mt h ln men dng lc y khng kh vi mt vng bn ngoi (outer loop) c pht trin cho ln men hiu kh i hi c s chuyn nhit. Mi trng v khng kh c a vo trong cc phn cao hn v thp hn (Hnh 4.14 c). Khng kh phc v cho hai mc ch: cung cp oxygen cn thit cho s sinh trng ca t bo v to ra s lu thng t nhin ca cht lng trong h ln men thng qua mt ci vng. B phn trao i nhit lm lnh mi trng lng c ci t vo trong ci vng . H ln men ny c chng minh l to ra mt tc hp th oxygen cao trn mt n v th tch.

Khng kh (a) (b) (c)

Hnh 4.14. Cc h ln men vng: (a) lc y khng kh, (b) lc y khng kh c bm bn ngoi, (c) p lc chu k ICI.

IV. Cc k hiu
B

tc chy ca dng x, m3/s t l x, c nh ngha nh B/F nng sn phm nng sn phm a vo nng c cht nng c cht im phi trn ca dng chy vo v dng chy thu hi

CP

CPi
CS

' S

Cng ngh t bo

55

C S0
CS f

nng c cht ti thi im t0 nng c cht sau khi ra khi h ln men nng c cht a vo

C Si
CX C
' X

CS ,opt nng c cht ti u

nng t bo nng t bo im phi trn ca dng chy vo v dng chy thu hi nng t bo ti thi im t0 nng t bo sau khi ra khi h ln men nng t bo a vo nng t bo thu hi qua lc nng t bo ca dng chy thu hi tc pha long, s-1 tc dng chy, m3/s thi gian lu, s

C X0
CX f

C Xi CX L CX R
D F

C X , opt nng t bo ti u

m,opt thi gian lu ti u, s p thi gian lu da trn tc dng chy ca ton b h thng

b
KS L

thi gian lu cn thit nui cy m hoc PFF trng thi n nh t ti mt nng t bo nht nh h s h thng tc dng chy qua lc, m3/s tc sinh trng c trng, s-1 hoc kg/m3/s tc sinh trng cc i tc thu hi tc sinh trng t bo th tch lm vic ca h ln men, m3 hiu sut sn phm/c cht

max
R rX V YP/S

Cng ngh t bo

56

YX/S X

hiu sut sinh trng/c cht t bo trn c s trng lng kh

Ti liu tham kho/c thm

1. Asenjo JA and Merchuk JC. 1995. Bioreactor System Design. Marcel Dekker, Inc. New York, USA. 2. Atkinson B and Mavituna F. 1991. Biochemical Engineering and Biotechnology Handbook. 2nd ed. Stockton Press, New York, USA. 3. Flickinger MC and Drew SW. 1999. Encyclopedia of Bioprocess Technology: Fermentation, Biocatalysis and Bioseparation. John Wiley & Sons, New York, USA. 4. Lee JM. 2001. Biochemical Engineering. Prentice Hall, Inc. USA. 5. Shuler ML and Kargi F. 2002. Bioprocess Engineering-Basic Concepts. 2 ed. Prentice Hall, Inc. NJ, USA.
nd

6. Vogel HC and Todaro CL. 1997. Fermentation and Biochemical Engineering Handbook (Principles, Process Design, and Equipment). 2nd ed. Noyes Publications. New Jersey, USA.

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57