Bai Tieu Luan Penixillin

8/8/2019 Bai Tieu Luan Penixillin

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I HC QUC GIA TP HCMTRNG I HC BCH KHOA

KHOA CNG NGH HA HC

Tiu lun mn hc

Cng ngh ln men ti :Nghin cu quy trnh sn xut Penicillin

GVHD : C Nguyn Thy HngLP :HC07BSHSVTH :


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Phn II : Ni dung

I.TNG QUAN V CHT KHNG SINH :S pht trin v vi sinh vt hc ni chung, v vi sinh vt cng nghip ni

ring, vi bc ngoc lch s l pht minh v i v cht khng sinh ca Alexander

Fleming (1928) m ra k nguyn mi trong y hc: khai sinh ra ngnh cng ngh

sn xut cht khng sinh v ng dng thuc khng sinh vo iu tr cho con ngi.

Thut ng "cht khng sinh" ln u tin c Pasteur v Joubert (1877) s

dng m t hin tng km hm kh nng gy bnh ca vi khun Bacillus

anthracis trn ng vt nhim bnh nu tim vo cc ng vt ny mt s loi vi

khun hiu kh lnh tnh khc.Nicolle (1907) l ngi u tin pht hin ra hot tnh khng khun ca

Bacillus subtilis c lin quan n qu trnh hnh thnh bo t ca loi trc khun ny.

Gratia v ng nghip (1925) tch c t nm mc mt ch phm c th s dng

iu tr hiu qu cc bnh truyn nhim trn da do cu khun.

Mc d vy, trong mi ti nm 1929 thut ng " Cht khng sinh" mi c

Alexander Fleming m t mt cch y v chnh thc trong bo co chi tit v

penicillin.

Thp k 40 v 50 ca th k XX ghi nhn nhng bc tin vt bc cangnh cng ngh sn xut khng sinh non tr, vi hnglot s kin nh :

-Khm ph ra hng lot cht khng sinh, th d nh Griseofulvin (1939), gramicidin S(1942) , Streptomycin (1943), bacitracin (1945), cloramphenicol v polymicin (1947),clotetracyclin v Cephalosporin (1948), neomycin (1949), oxytetracyclin v nystatin(1950), erythromycin (1952), cycloserin (1954), amphotericin B v Vancomycin(1956), metronidazol, kanamycin v rifamycin (1957)...

-p dng phi hp cc k thut tuyn chn v to ging tin tin (c bit l cc k

thut gy t bin, k thut dung hp t bo, k thut ti t hp gen ...) to ra

nhng bin chng cng nghip c nng lc "siu tng hp" cc cht khng sinh caogp hng vn ln cc chng ban u.

-Trin khai thnh cng cng ngh ln men chm quy m sn xut cng nghip sn

xut Penicillin G (1942) v vic hon thin cng ngh ln men ny trn cc sn

phm khc.


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-Vic pht hin, tinh ch v s dng axit 6 - aminopenicillanic (6-APA, 1959) lm

nguyn liu sn xut cc cht khng sinh penicilin bn tng hp cho php to ra

hng lot dn xut penicilin v mt s khng sinh - lactam bn tng hp khc.

1.Khi nim cht khng sinh:

Khng sinh l nhng cht c ngun gc vi sinh vt, bn tng hp hay tng hp.Vi liu iu tr khng sinh c tc dng km hm hoc tiu dit vi sinh vt gy bnhngay nng thp. Mt s khng sinh c tc dng ngn cn s pht trin ca t boung th.

So vi thuc st khun, thuc khng sinh t c vi c th hn v khng sinh ckh nng c ch chn lc i vi mt s khu trong qu trnh pht trin ca vi khungy bnh. Tuy nhin khng sinh khng phi l cht v hi i vi c th, mt skhng sinh c th gy c vi gan thn , h thng to mu hoc gy ri lon tiuha.

2. C ch tc dng:C ch tc dng ln vi sinh vt gy bnh ( hay cc i tng gy bnh khc - gi

tt l mm bnh) ca mi cht khng sinh thng mang c im ring, ty thuc vo

bn cht ca khng sinh ; trong , nhng kiu tc ng thng gp l lm ri lon

cu trc thnh t bo, ri lon chc nng iu tit qu trnh vn chuyn vt cht ca

mng t bo cht, lm ri lon hay kim to qu trnh sinh tng hp protein, ri lon

qu trnh ti bn ADN, hoc tng tc c hiu vi nhng giai on nht nh trong

cc chuyn ha trao i cht (hnh 1)

Hnh 1. V tr tc dng chnh ca mt s cht khng sinh


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3. n v khng sinh:

Nng lc tch t khng sinh ca chng hay nng cht khng sinh thng

c biu th bng mt trong cc n v l : mg/ml, g/ml, hay n v khng sinh

UI/ml

(hay UI/g,International Unit).

n v ca khng sinh c nh ngha l lng khng sinh ti thiu pha

trong mt th tch quy c dung dch c kh nng c ch hon ton s pht trin ca

chng vi sinh vt kim nh chn ; th d, vi penicillin l s miligam penicillin

pha vo trong 50 ml mi trng v s dng Staphylococcus aureus 209Plm chng

kim nh .

4. Hot tnh khng sinh c hiu:

Hot tnh khng sinh c hiu l c tnh cho thy nng lc km hm hay tiu

dit mt cch chn lc cc chng vi sinh gy bnh, trong khi khng gy ra cc hiu

ng ph qu ngng cho php trn ngi bnh c iu tr. c tnh ny c biu

th qua hai gi tr:

Nng km hm ti thiu (Minimun Inhibitory Concentration - Vit tt l MIC) v

nng dit khun ti thiu (Minimun Bactericidal Concentration - Vit tt l

MBC), xc nh trn cc i tng vi sinh vt gy bnh kim nh la chn tng

ng cho mi cht khng sinh.

5.Hin tng khng thuc :

5.1 Hin tng khng thuc: Hin tng mm bnh vn cn sng st sau khi iu

tr khng sinh c gi l hin tng khng thuc (trn phng din kim nghim, vi

sinh vt gy bnh c coi l khng thuc nu nng MIC ca cht khng sinh

kim nghim in vitro trn i tng ny cao hn nng iu tr ti a cho php i

vi bnh nhn.

5.2 Nguyn nhn hin tng khng thuc:

- Vic s dng cng loi khng sinh ko di hoc lm dng thuc khng sinh (tu tin

s dng thuc khng ng liu lng, khng ng ch nh v khng thi gian cn

thit) v tnh to ra u th pht trin cnh tranh cho cc chng vi sinh vt c khnng khng thuc .

- Xu th s dng tu tin cht khng sinh trong chn nui, c bit l b sung vo khi

ch bin thc n gia sc, gia cm nui ly tht, trng, sa ... Khi , ngoi cc tc

dng c li d kin, chnh cht khng sinh b sung s to ra mi trng pht trin

chn lc cho cc chng mang yu t khng thuc R trn ng vt nui. Khi s dng


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tht, trng, sa ... ca chng lm nguyn liu ch bin, cc chng khng thuc ny s

ko theo vo trong cc sn phm thc phm. Kt qu khi ngi tiu dng s dng cc

thc phm ny, mt mt h phi tip nhn phn d lng khng sinh trong sn phm;

nhng mt khc, nguy him hn l cc loi vi sinh vt khng thuc trong cc sn

phm thc phm thuc nhm ny c u th tn ti, pht trin cao hn .

5.3 Cch khc phc :

Khc phc hin tng khng thuc ca vi sinh vt gy bnh: gii php trc quan v

n gin l s dng cc dng khng sinh mi. Tuy nhin, vic tm kim, pht hin v

sn xut mt khng sinh mi l c mt khi lng cng vic khng l, tiu tn rt

nhiu thi gian, nhn lc v tin bc

Trc ht cn trit tn trng ba nguyn tc s dng thuc khng sinh l:

-Ch nh iu tr khng sinh ng (lm khng sinh chn ng khng sinh thch

hp ch nh iu tr; dng thuc ng liu, ng phc , thi gian iu tr;

ch pht hin sm du hiu khng thuc);

-Khng lm dng khng sinh khi cha cn thit (khng lm dng "iu tr phng

nga" bng thuc khng sinh, nghim cm bnh nhn t ch nh iu tr thuc khng

sinh thay bc s);

-Nghim cm s dng trn lan cht khng sinh trong chn nui v gim st cht ch

vic s dng khng sinh trong th y.

II .CHT KHNG SINH PENICILLIN :

1.Lch s pht hin v sn xut Penixillin :

Ngi u tin pht hin ra khng sinh l A. Fleming. Nm 1928, khi nghin cu cc

vi sinh vt gy bnh , ng tnh c pht hin ra VSV gy bnh b tiu dit bi mt loi

nm si mu xanh xm , ng trch li dch ny nh ln khun lc ca cc vi khun gy

bnh, kt qu l VSV b cht.

Nm to nn cht tiu dit vi sinh vt l Penicillium. Cht do nm tit ra gi l

Penicillin. Mt nm sau ng cng b kt qu nghin cu nhng khng c quan tmn.

n nm 1938, Howard Florey, Ernst Chain, Norman Heatley mi a Penicillin vo

sn xut th. Vo nm 1940, Dorothy Hodkin xc nh c cu trc phn t ca

Penicillin.


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1942: Mary Hunt tuyn chn c chng cng nghip Penicillium chrysogenum

NRRL 1951 (1943) v sau c bin chng P. chrysogenum Wis Q - 176

(chng ny c xem l chng gc ca hu ht cc chng cng nghip ang s dng

hin nay trn ton th gii) thnh cng trong vic iu chnh ng hng qu

trnh ln men ln men sn xut penicillin G (bng s dng tin cht Phenylacetic,1944)....

Nm 1959, cc nh khoa hc Anh v M tch ra c axit 6-aminopenicillanic.

y l nguyn liu sn xut ra hng lot ch phm penicillin bn tng hp khc

nhau. Ngy nay trn th gii sn xut ra c trn 500 ch phm penicillin ( trong

ch ln men trc tip hai sn phm l penicillin V v penicillin G) v tip tc trin

khai sn xut cc ch phm penicillin bn tng hp khc.

Tnh hnh sn xut penicillin hin nay trn th gii :

Thuc khng sinh chim khoang 30% th phn dc phm th gii. Cc tp ondc phm ca M, Anh, Nht Bn , Italia, H Lan, n chim cc v tr hang

u trong lnh vc ny. V d : Cng ty DSM c tr s ti thnh ph Delf (H Lan)

sn lng hng nm 15000 tn. Penicillin chim 30% sn lng loi thuc ny trn

th gii

-Lch s tuyn chn chng cng nghip P. chrysogenum

Vo nhng nm u, vic nghin cu

sn xut penicillin thng s dng cc

chng c hot lc cao thuc loi P. notatum

v P. baculatum. Nhng t khi trng i

hc Wisconsin (M)phn lp c chng

P.chrysogenum c hot tnh cao hn th

chng ny dn dn thay th v t khong


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sau nhng nm 50 ca th k XX n nay tt c cc cng ty sn xut penicillin trn

th gii u s dng cc bin chngP.chrysogenum cng nghip.

Vic tuyn chn chng cng nghip ln men sn xut penicillin trn nguyn

tc cng tri qua su giai on c bn, trong gii php k thut c p dnghiu qu thu nhn bin chng "siu tng hp" penicillin li chnh l cc k thut

gy t bin thng nh: x l tia Rn - ghen, x l tia cc tm v to t bin bng

ho cht, th d nh Metylbis amin(metyl -2- -clo- etylamin), N-mustar (tris - -

clo- etylamin), Sarcrolyzin, HNO2, Dimetylsulfat, 1,2,3,4 -diepoxybutan.

3.Tnh cht ha l ca penixillin :

Tnh cht vt l:

Cc penicillin di dng mui hoc dng acid l nhng bt trng khng mikhi tinh khit.

Ph UV: a s cc nhm R acyl ha trn 6-APA u l vng thm nn cho

ph hp thu vng UV c th ng dng c.

Ph IR: vng 1600 1800 cm-1 c cc nh c trng cc nhm sau :

-Nhm lactam gia 1760 v 1730 cm-1.

-Chc carboxyl khong gia 1600 cm-1.

-Nhm chc amid ngoi vng gia 1700 v 1650 cm-1.

Bnh thng penicillin c trng lng phn t ca 313 n 334 g / mol ( penicillinG). Penicillin vi cc nhm phn t nh km thm c th c mt khi lng phn tkhong 500 g / mol. V d, cloxacillin c khi lng mol ca 476 g / mol vdicloxacillin c khi lng mol ca 492 g / mol.

Tnh cht ha hc:

Cc penicillin c kh nng to mui natri v kali tan trong nc, trong khi

cc mui kim loi nng ( vi d mui Cu2+) th khng tan hoc kch thch sphn hy.

Cc penicillin cng c kh nng to mui vi cc amin:
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-To cc penicillin thy gii chm ( tc ng tr) nh procain penicillin

( tc ng ko di t 24 48h), benethamin penicillin ( tc ng ko di

th 3 7 ngy), benzathin penicillin ( tc ng ko di 2 4 tun).

-Mt s c tnh base v d cc aminosid, cc alkaloid khi trn chung vipenicillin trong cng mt ng tim s gy ra kt ta.

-Cc penicillin cng c kh nng to ra cc este, s l nhng tin cht c

kh nng phng thch cc khng sinh ny trong invivo.

Tnh khng bn ca vng beta lactam:

S phn hy trong mi trng kim: pH = 8 s c s tn cng ca ion

OH- trn carbonyl lactam gy ra s m vng theo qui lut chung, cui cng s

c s to thnh acid penicilloic, nhng s decarboxyl c th xy ra tip theo

to acid peniloic.

Nu trong mi trng c s hin din ca nhng mui kim loi nng ( Zn2+,

Cd2+, Pb2+ hoc Hg2+) s lm cho acid peniciloic b phn hy thnh

carbinolamin khng bn, cht ny s tip tc b phn hy to D-penicillamin v

acid penaldic.Acid penaloic n lt n c th b decarboxyl ha tr thnh

penicillo-aldehyd.

S alcol phn v amino phn: vng beta lactam nhy vi mt s tc nhn i

nhn khc vi xc tc ca cc ion kim loi nng: Cu2+, Zn2+, Sn2+.

S phn hy trong mi trng acid: di s hin din ca ion H +, s tn

cng i in t trn nguyn t S, kch thch s m vng lactam v vng

thiazolidin, tip theo l s ti sp xp to thnh cu trc oxazolic cua acid

penicillenic. Cui cng, nu mi trng qu acid c th to thnh acid penillic.

Ngoi ra vng lactam c th b m bi lactamase tit ra t vi khun.

2. Phn loi Penixillin :

- c dim chung :

Penixillin l dn xut ca 6-aminopenixilamic gm 1 vng thiazolidin v mt vng

beta-lactam.


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-Cng thc chung ca penixillin :

- -

-

--

-

- Khi thay th H bng kim loi kim hoc kim th s c cc penixillin d

tan trong nc (nh Na, K, Ca )

- Vng beta-lactam l yu t quyt nh hot tnh ca khng sinh

- Khi thay th R bng cc gc khc nhau s c cc penixillin c tc dng

khc nhau

Da vo ngun gc c th sp xp penixillin vo 3 nhm :

-Penixillin nhm I :gm cc penixillin t nhin c chit xut t mi trng nui

cy nm penixillium notatum hoc P chrysogenum nh penixillin G hoc penixillin V

Cc penixillin c hp thu nhanh v thi tr ra khi c th nhanh nn thi gian tc

dng ngn. Mun ko di tc dng phi dung cc dn xut ca chng nh procain

benzyl penixillin (ko di trong 24 h) hoc benzathin benzyl penixillin ( ko di trong

4 tun )

Cc penixillin chm ch dung tim bp khng c tim tnh mch

-Penixillin nhm II : Gm cc dn xut ca cc penixillin bn tng hp c ph khng

khun hp hn Penixillin G nhng c kh nng khng penixilinase dng cha bnh

nhim khun do t cu khng penixillin nhm I nh methycylin, cloxaxilin


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- Penixilin nhm III : Gm cc penixillin bn tng hp ph rng, khng khng c

penixilase nhung khng c cc vi khun Gram(-) m cc penixillin nhm II t tc

dng bn vng trong mi trng dch v nn c th dung ung nh ampixillin,

amoxycilin.


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4.C ch sinh tng hp penicillin nm mc P. chrysogenum :

Nhn chung, c tng cng ba bc quan trng bc quan trng trong qu trnh sinhtng hp penicillin G (benzylpenicillin)

Bc u tin trong sinh tng hp ca penicillin G l ngng t ca ba axit amin L--aminoadipic acid, L-cysteine, L-valine thnh mt tripeptide .Trc khi ngng tthnh mt tripeptide, cc amino acid L-valine s tri qua qu trnh epimerization vtr thnh D-valine. Sau qu trnh ngng t , c t tn tripeptide -(L--aminoadipyl)-L-cysteine-D-valine (vit tt l ACV). Trong qu trnh phn ng cn 1enzyme xc tc l ACVs enzyme . ACVs enzyme ny cn thit cho vic kch hotca ba axit amin trc khi ngng t v epimerization ca bin i L-valine thnhvaline-D.

Bc th hai trong sinh tng hp ca penicillin G l s dng mt loi enzyme thay i ACV thnh isopenicillin N. Cc enzyme l synthase N isopenicillin vi genpcbC km theo. Cc tripeptide trn ACV s tri qua qu trnh oxy ha, sau khpvng . Isopenicillin N l mt trung gian rt yu bi v n khng hin th nhiu hotng khng sinh.

Bc cui cng trong sinh tng hp ca penicillin G l trao i cc nhm bnchui isopenicillin N s tr thnh penicillin G. Thng qua cc coenzyme xc tcisopenicillin N acyltransferase (IAT), pha alpha-aminoadipyl-chui isopenicillin Nc ly ra v trao i cho mt chui phenylacetyl-side. Phn ng ny c m hobi cc penDE gen. xem s tng hp penicillin G trong hnh 3.
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Hnh 3. C ch sinh tng hp penicillin t axit L- - aminoadipic, L-cystein v L-valin

5.Tc ng ca cc thng s cng ngh n qu trnh sinh tng hp

penicillin.

5.1. S pht trin h si v c im hnh thi h si nm:

S pht trin h si nm trong qu trnh ln men bao gm:

- S tng trng v kch thc h si (tng di si, s ln ln v kchthc, mc phn nhnh ca h si ... )

- S bin thin v s lng khm si nm trong mi trng: Thng thng, spht trin ny c nh gi qua hai ch tiu l: hm lng sinh khi v tc binthin hm lng sinh khi trong mi trng. Hai ch tiu ny c th xc nh bngnhiu phng php khc nhau nh: hm lng sinh khi (Sinh khi ti hoc sinhkhi kh), mt quang dch ln men, tr lc lc ca dch ln men, hm lng nit,hm lng hydratcacbon, hm lng axit nucleic ... Trong cc phng php trn,c p dng ph bin hn c trong sn xut cng nghip l phng php xc nhqua hm lng sinh khi.

Tc pht trin h si nm ph thuc hng lot cc yu t khc nhau trong qutrnh ln men v s tch t penicillin thng xy ra mnh m khi h si pht trin ttrng thi cn bng. Trng thi ny c th xc lp c khi ch cung cp va vlin tc lng thc n ti thiu cho nm mc. Thiu thc n, h si nm s t phn,cn nu cung cp qu nhu cu trn, h si s pht trin, nhng khng tch t mnhpenicillin m tch t nhiu axit gluconic v axit malic.

-c im hnh thi v cu trc h si nm: Trong qu trnh ln men, do

nhiu nguyn nhn khc nhau, s lng khm si nm bao gi cng c xu hng


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tng ln, ngay c trong qu trnh ln men tnh. Trong iu kin ln men c sc kh v

khuy trn, do tc dng va p c hc vi cnh khuy v cc chuyn ng dng xoy

trong mi trng, mt mt s t gy h si nm xy ra nhiu hn v h si nm bao

gi cng c xu hng vn cun li thnh cu trc bi si cun xon, c gi l

pellet.

Pellet xp (fluffy loose pellets) l dng pellet c phn bn trong h si cun thnh

khi chc v mn, lp si pha bn ngoi cun lng lo to thnh cu trc xp hn.

Pellet chc v mn (compact smooth pellets) c c im l phn si pha bn trong

pellet cun tng i cht ch ra n gn st lp si pha ngoi, lp si pha ngoi

cng cng cun chc thnh lp si mn.

Pellet rng (hollow pellets) l dng pellet c phn si bn trong b t phn to thnh

khong rng, h si pha bn ngoi cun rt cht thnh lp si mn v chc chn.

- Hiu qu chung ca qu trnh ln men c quan h hu c vi s lng, kch thc

v cu trc pellet nm. Trong thc tin sn xut cng nghip, ngi ta thng iu

chnh cc thng s cng ngh theo hng u tin to ra dng pellet nh v mn,

hn ch to pellet xp v ngn nga hnh thnh cc pellet rng. iu kin cng ngh

tng ng vi mc tiu trn thng p dng l : t l cy ging 10%, vi mt dch

ging

(2-10).1011 bo t /m3; phi hp iu chnh gia sc kh v khuy trn m bo

cung cp oxy ha tan d so vi nhu cu tng ng vi thi im ln men, v to ra

pellet mn v nh (kch thc pellet thch hp nht khong 0,2 - 0,5mm), trong iukin cn i vi nhu cu tit kim mc tiu tn nng lng do khuy trn.

5.2. c tnh nhit ng ca dch ln men:

Trong cc thit b ln mendung tch ln c sc kh v khut trn, thc t khng th

xc lp c s ng u ti khp cc vng th tch lm vic ca thit b. Ti cc

vng chy ri (vng gn cnh khuy), tc trao i nhit, tc chuyn khi xy ra

mnh m hn. Cn ti cc vng chy mng (vng st thnh thit b, vng gn cc ng

xon trao i nhit, vng km hiu qu hay vng cht ca thit b) tc chuyn

khi hay tc truyn nhit cng gim i. Ngoi ra, ti nhng khu vc nht nh cathit b c th xut hin vng xoy cc b hay cc dng chy th cp lm thiu ht v

hm lng oxy ha tan.

Cc yu t nu trn y s tc ng trc tip n nng lc sinh tng hp ca chng,

hiu qu chuyn ha to sn phm v hiu qu kinh t chung ca ton qu trnh ln

men. Thc t thng chn ch khuy trn d trn mc yu cu.


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5.3. Thnh phn mi trng ln men:

Mi trng c s ln men penicillin, vo thi k u trong nhng nm 40 - 50, l

mi trng lactoza - nc chit ng

Ngun c cht chnh: l lactoza c th c thay th tng phn hoc ton b bng cc

c cht khc nh: cc loi ng hexoza, ng pentoza, disaccarit, dextrin hay thay

th bng du thc vt. Trong cc c cht nu trn, hiu qu cao hn c vn l glucoza.

Ngoi ra, khi s dng du thc vt lm cht ph bt phi xt n hiu ng nm mcs dng mt phn du thc vt lm ngun cung cp thc n cacbon, tnh ton iuchnh nng glucoza trong mi trng ln men (v c s cn tr qu trnh chuynkhi do nh hng ca du ph bt).

Ngun cung cp thc n nit: c th s dng l bt u tng, bt ht bng, cc loidu cm. Nhu cu v thc n nit cng c th c p ng bng cch cung cp lintc (NH4)2SO4, nhng duy tr nng thp, khong 250 - 340g/l (nu d tha hiu

qu sinh tng hp penicillin s gim, nu thiu s xy ra hin tng t phn h si) .Hm lng cc cht khong b sung: c tnh ton, ph thuc vo lng dch chitng s dng; pH mi trng c iu chnh trc khi thanh trng, sau trong sutqu trnh ln men c gim st cht ch v iu chnh theo yu cu cng ngh.

Nng tin cht to nhnh:Trong qu trnh sinh tng hp penicillin, vic kt gnmch nhnh ca phn t penicillin khng mang tnh c hiu cht ch. Nh vy, nuduy tr nng tin cht to nhnh cn thit phenylacetat (hoc phenooxyacetat) scho php thu nhn ch yu mt loi penicillin G trong dch ln men (hoc penicillinV). Theo l thuyt, nhu cu v phenylaceta l 0,47g/gam penicillin G (hocphenooxyacetat l 0,50g/gam penicillin V ). Cn ch c hai cu t trn thc cht

u gy c cho nm nn ngi ta thng la chn gii php b sung lin tc cu tny v khng ch cht ch nng theo yu cu, khng lm suy gim nng lc lnmen ca chng sn xut.

5.4. iu kin tin hnh ln men:

Nhit l thng s c nh hng ln n s pht trin ca nm mc, kh nng sinh

tng hp v nng lc tch t penicillin ca chng. Nhn chung nm mc pht trin

thun li hn di nhit khong 300C. Tuy nhin, di nhit ny tc phn

hu penicillin cng xy ra mnh m. Trong thc t, giai on nhn ging sn xut

ngi ta thng nhn di nhit 300C; sang giai on ln men thng p dngmt trong hai ch nhit l :

Ln men mt di nhit : Thng duy tr nhit trong sut qu trnh ln men

di nhit 25 - 270C.

Ln men hai ch nhit : Giai on ln men bt u tin hnh 300C cho n

khi h si pht trin t yu cu v hm lng sinh khi th iu chnh nhit sang


15/37

ch ln men penicillin di nhit 22 - 250C (c cng ngh iu chnh xung 22

- 230C, gi nhit ny tip hai ngy ri chuyn sang ln men tip 25 0C cho n

khi kt thc qu trnh ln men).

pH mi trngthun li cho s pht trin h si v cho qu trnh sinh tng hp

penicillin thng dao ng trong khong pH = 6,8 - 7,4. Tuy nhin iu kin pH

cao xu hng phn hu penicillin cng tng ln. V vy, trong sn xut pH mi

trng thng c khng ch cht ch gi tr la chn trong khong pH = 6,2 -

6,8.

Nng oxy ho tan v cng khuy trn dch ln men: Vi nhiu chng nm

mc, nng oxy ha tan thun li cho qu trnh sinh tng hp penicillin dao ng

quanh mc 30% nng oxy bo ha.

Nng CO2 trong dch ln men mc nht nh cng cn thit cho qu trnh ny

mm ca bo t nm mc; tuy nhin nu nng CO2 qu cao s lm cn tr qutrnh hp thu v chuyn ho c cht ca chng, ngha lm lm cn tr qu trnh sinh

tng hp penicillin.

5.5. S tch t v phn hu penicillin:

Trong qu trnh ln men, do nhiu nguyn nhn khc nhau, trong c nh hng

ca nng penicillin tch t trong mi trng ngy cng tng, lm cho nng lc sinh

tng hp penicillin ca chng c xu hng gim dn theo thi gian ln men. ng

thi, ph thuc vo nhit v pH mi trng, mt phn lng penicillin tch t cng

b phn hu theo thi gian.Nhm gim tn tht trn, ngay sau khi kt thc qu trnh ln men cn x l thu sn

phm sm hoc c gii php h thp nhanh nhit dch ln men.

III .QUY TRNH LEN MEN SN XUT PENIXILLN TRONG

CNG NGHIP :

1. c im chung:

Nhng vi sinh vt sinh penicillin thuc cc ging nm mc penicilliumv Aspergillus. Nhng cc chng thuc nhm Penicillium notatum, Penicillium

chrysogenum c hot lc cao v c dng trong cng nghip khng sinh.

Nhng chng u tin c nui cy bng phng php b mt trn c s cht

t nhin to thnh 10-15 v/ml khng sinh.


16/37

Penicillium chrysogenum trn mi trng Raistrik to thnh hai kiu

khun lc:

-Kiu I: khun lc trn trn, cc np nhn r nt. Khun ty kh sinh mc tt v

c mu xanh; theo ra khun lc c ng vin rng 2-5 mm ca nhng khunty bc trng khng c bo t; cc khun ty c cht mu nu; cht mu khng

ho vo mi trng.

-Kiu II: khun lc c nhng khun ty mu trng, pht trin yu; khun ty c

cht cng c mu nu. Khun lc kiu I cho hot l cao, kiu II thng xuyn

cho hot tnh khng sinh thp. V vy cn phi tch nhng khun lc kiu I trn

mi trng ny v thng xuyn kim tra chn nhng khun lc c hot lc

cao, gi c c tnh ca ging.

Chng penicillium c nui cy trn a petri


17/37

Cc chng penicillium cc giai on pht trin khc nhau

Nhng chng Penicillium thng c hot lc cao li km n nh. c

tnh ny t cho nhng nh vi sinh vt mt nhim v kh khn: to c kh

nng sinh khng sinh cao nht, gi c n nh trong qu trnh nghin cu v

sn xut. Nhim v ny c mt ngha rt ln trong cng nghip .cc ging

ny bo v k, trng thi ng kh c th ti ba nm, t v trng l hai

nm. Ngy nay nh di truyn hc to c nhng ging n nh, t nht sau

su th h khng gim hot tnh khng sinh.

Chng ta cn phi nhn thc rng cc nm penicillium thng d bin

i v hnh thi v gim kh nng sinh khng sinh. Khi xy ra bin i th s

sinh ra hng lot nhng chng mi t ging c bn v nhim v ca cc nh vi

sinh vt lc ny l phi chn li nhng khun lc kho c nhiu u im, tip

theo cn phi tin hnh nhng bin php bo qun thch hp.

Trong qu trnh nui cy chm nm Penicillium chrysogenum tri qua su giai on

pht trin:

- Giai on I: Cc bo t nm mc ny mm, pht trin thnh chi nh , t bo cht

cha phn ha. Thnh thong khng bo c nhng ht nh bt mu trung tnh.


18/37

- Giai on II: Khun ty pht trin, t bo cht a kim, nhng ht nh trong khng

bo dn dn bin mt. cui giai an ny xut hin nhng git cht bo nh .

- Giai on III: To thnh nhng git cht bo to, khng cn khng bo, t bo cht

rt a kim.- Giai on IV : Xut hin khng bo vi nhng ht d bt mu trung tnh, nhng

ht cht bo nh hn giai on III, tnh a kim gim

- Giai on V : Khun ty c hnh trng v c nhng khng bo, giat hoc vi ht

ln . Cc ht cht bo bin mt , tnh a kim gim

- Giai on VI : Khun ty vn c dng hnh trng nhng khng cn nhng ht bt

mu trung tnh, khng bo bt mu da cam, hoc mu hng ng u. Cc ht cht

bo khng cn. Xut hin nhng t bo ring bit bt u t phnQu trnh ln men penicillin cng thuc loi ln men 2 pha : Pha sinh trng ng vi

giai on I,II,III . Pha sinh penicillin ng vi giai on IV,V,VI

Cng ngh ln men sn xut penicillin mang nt c th ring ca tng c s sn xut

v cc thng tin ny rt hn ch cung cp cng khai, ngay mi bng sng ch thng

cng ch gii hn nhng cng on nht nh; v vy rt kh a ra c cng ngh

tng qut chung. Theo cng ngh ln men ca hng Gist-Brocades (H Lan), ton b

dy chuyn sn xut thuc khng sinh penicillin c th phn chia lm bn cng on

chnh nh sau (xem s hnh 4)

Ln men sn xut penicillin t nhin (thng thu penicillin V hoc penicillin G) .

X l dch ln men tinh ch thu bn thnh phm penicillin t nhin.

Sn xut cc penicillin bn tng hp (t nguyn liu penicillin t nhin)


19/37

Pha ch cc loi thuc khng sinh penicillin thng mi.

Hnh 4. S dy chuyn sn xut penicillin

(theo Gist-Brocades Copr. (H Lan))

- S khi :


20/37

Chun b ln men :

Ging, bo qun v nhn ging cho sn xut:

Ging dng ln men penicillin l P.chrysogenum, y l loi nm si bo t h.

Khi mi pht hin trong mi trng c, chng to ra hai loi khun ty: khun ty khsinh v khun ty dinh dng mu trng. Sau khi nui cy 1 ngy, khun ty bt uchuyn sang mu xanh xm v nh bo t bt u xut hin . Thi gian ny bt uxut hin 1 t bo t trn t tin bo t nm trong cc nh bo t. Cc bo t ln lcc to thnh theo thi gian nui cy v cui cng th mu ca nm penicillum smhn

-Ging cng nghipP.chrysogenum c bo qun lu di dng ng kh, bo qun

siu lnh -70oC hoc bo qun trong nit lng. Ging t mi trng bo qun c


21/37

cy chuyn ra trn mi trng thch hp hot ho v nui thu bo t. Dch huyn

ph bo t thu t hp petri c cy chuyn tip sang mi trng bnh tam gic, ri

sang thit b phn ging nh, qua thit b nhn ging trung gian ... v cui cng l

trn thit b nhn ging sn xut. Yu cu quan trng ca ca cng on nhn ging

l phi m bo cung cp lng ging cn thit, vi hot lc cao, cht lng mbo ng thi im hco cc cng on nhn ging k tip v cui cng l cung cp

lng ging t cc yu cu k thut cho ln men sn xut. Trong thc tin,

m bo cho qu trnh ln men thun li ngi ta thng tnh ton lng ging cp

sao cho mt ging trong dch ln men ban u khong 1 - 5.109 bo t / m3.

Thnh phn mi trng nhn ging cn c tnh ton m bo cung cp

ngun thc n C, N, cc cht khong v cc thnh phn khc, m bo cho s hnh

thnh v pht trin thun li ca pellet.

Chun b mi trng ln men v thit b:- Chun b mi trng ln men:

. Cn ong, pha ch ring r cc thnh phn mi trng ln men trong cc thngcha ph hp

. Thanh trng gin on 1210C ( hay thanh trng lin tc khong 140-1460C)hoc lc qua cc vt liu siu lc ri mi bm vo thng ln men.

Nu c tnh cng ngh ca thit b ln men cho php, c th pha ch ri thanhtrng ng thi dch ln men trong cng mt thit b. Tt c cc cu t b sung vomi trng ln men u phi c x l kh khun trc v sau b sung theo ch

vn hnh v khun.Thit b ln men: Phi c v khun trc khi a vo s dng. Thng thanh

trng bng hi qu nhit 2,5 3,0 at trong thi gian 3 gi. ng thi kh khun

nghim ngt tt c cc h thng ng dn, khp ni, van, phin lc v tt c cc thit b

ph tr khc.Trong qu trnh ln men luun c gng duy tr p sut d trong thit

b nhm hn ch ri ro do nhim tp.

Khng kh thng c kh khun s b bng nn on nhit, sau qua mng

lc v khun hay mng siu lc .

3. K thut ln men:3.1. K thut ln men b mt:

Trong nhng nm 30 n nhng nm 50 ca th k XX, phng php

nui cy b mt c p dng rng ri sn xut khng sinh t nm


22/37

penicillin chrysogenum ngy nay k thut len6men b mt c thay th

bng k thut ln men chim

a) GingPenicillin chrysogenum: l nm si n bo h. Khi mi pht trin trong

mi trng t th to ra 2 khun ty: khun ty kh sinh v khun ty dinh dng mutrng. Sau 1 ngy nui cy, cc khun ty chuyn sang mu xanh sm v nh bo t

bt u xut hin. Thi gian ny cng xut hin mt t bo t trn t tin bo t nm

trong cc nh bo t. Cc bo t ln lt to thnh theo thi gian nui cy v cui

cng nm penicillin c mu sm hn.

Nm si penicillium chrysogenum.

Nm penicillin: thuc h hiu kh bt buc, do trong qu trnh nui phi

cung cp kh lin tc. Phng php bo qun c dng nhiu nht l phng phpcy truyn nh k trn thch hng thng kt hp vi bo qun lnh, phng php bo

qun bng ht ng cc bo qun theo phng php ng kh cng c s dng.

b)Nguyn liu: Cm v ht ng cc cc lai, nguyn liu c b sung nc sao cho

m t 55-60%W v c hp thanh trng 121oC trong 30-45 pht. Ngay sau

khi kt thc thanh trng, chng c ti vo nhng khay hnh ch nht c kch thc

di 1-1.2 m, rng 0.6-0.8 m, cao 5-6 cm. Lp mi trng cho vo y dy 2-3 cm

m bo thang kh trn tan b b mt v mt di ca mi trng. Mt s c sdng nguyn liu l cc ht ng cc th lp mi trng dy hn (3-4 cm) do cc ht

ng cc to ra mi trng c thong kh hn.

Trong trng hp cm qu mn th phi trn thm tru xay nh (thm khong

20-25%) hoc ci bp xay nh trc khi thanh trng.


23/37

lm mi trng nhn ging, ngi ta cng lm nh trn. Ch c mt im

khc l sau khi lm m mi trng n m nht nh, ngi ta phn phi chng

vo cc dng c thy tinh (chai thy tinh hay bnh tam gic) vi khi lng bng 1/5

hay 1/6 dung tch ca dng c, y kn bng nt bng v thanh trng 121

o

C(0.5 at)trong 30 pht ri ngui mi cy ging.

c) Qu trnh nhn ging: bt u t ging c trong ng nghim. Trong cc nh my,

mi ln cy truyn ging, ngi ta thng cy lm 3 ng. Mt ng dng kim tra

trc khi sn xut, mt dng sn xut v mt dng bo qun.

Song song , ngi ta chun b mt bnh tam gic dung tch 200-250ml v chun b

50g mi trng. Mi trng c thanh trng v lm ngui n 30oC.

10ml thanh trng v lm ngui vo ng ging, dng que thy tinh nh

cho bo t ha trn vo nc. Bng bin php v trng (thc hin trong cc t cy v

trng) chuyn ton b vo bnh tam gic trn, lc cho tht u ri chuyn chng sang

t m30-37oc. Nui iu kin ny cho n khi bo t nm xut hin v pht trin

u khp mi trng.

Ta gi qu trnh thc hin nh trn l qu trnh nhn ging cp 1.C tip tc

thc hin ta c ging cp 2, cp 3 cho n khi 5-10% ging cho sn xut.

C mi mt cp nhn ging t cp ny sang cp khc, khi lng mi

trng tng t 10-15 ln. Trong trng hp vt qu 1 k ngi ta nui trn nhngkhay.

d)Qu trnh ln men: khi mi trng c kh trng v lm ngui n 30oC, tin

hnh trn ging vi t l 5-10%. Cc khay c xp chng ln nhau trn nhng gi

vi khong cch nht nh thong kh v thong nhit.

Nm penicillium trong qu trnh pht trin thng to t nhit hn nm

Aspergillus. Tuy nhin, tng cng kh nng pht trin v sinh tng hp, ngi ta

thng thi kh bng qut gi c lp h thng lm sch.

Qu trnh ln men ko di 6-7 ngy 24-28oC.

Trong ln men b mt, ngi ta s dng mi trng lng. Mi trng lng

dng trong nui cy b mt thu nhn penicillin bao gm:

Cao ng (bp) 50g Lactose 30g NaNO3 3gKH2PO4 0.5g MgSO4 0.25gC6H5CH2COOH 0.2g ZnSO4 0.044g Nc 1000ml


24/37

Dung dch ln men c kh trng 121oC (0.5 at) trong 30 pht, c phn

khi vo cc khay ging cc khay nui cy b mt vi mi trng bn rn. y cc

khay ny khng c c l v phi cha mi trng lng. Chiu cao ca dung dch

mi trng trong cc khay l 3-4 cm. Ngi ta cng tin hnh ln men trong khong

thi gian l 6-7 ngy nhit ln men l 24-28oC.

Vng nm si c gi li sau khi rt ht dch ln men, c tip tc s

dng cho nhng ln ln men k tip. nhng ln ln mentip theo ngi ta ch

thm dch ln men vo. Cc th nghim cho thy ch nn s dng li 3-4 ln, v nhng

ln sau hiu sut thu nhn khng sinh s gim dn.

Khu vc ln men sn xut penicillin.

3.2. K thut ln men chm:

K thut len men chm l k thut c p dng trong hu ht cc c c sn xut

penicillin cng nghip hin nay v thng c vn hnh theo phng php ln menbn lin tc, gm phng n ln men gin on theo m c b sung lin tc (hay bn

lin tc) mt hay mt vi cu t kt hp vi phng n tun hon li mt phn h si

ca m ln men trc (hoc khng)


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3.2.1 Qu trnh nhn ging :

Trong qu trnh ln men chm ngi ta nhn ging trong mi trng lng. Mc ch ca

qu trnh nhn ging l thu nhn c s lng t bo cao ( thng tnh tng lng t

bo/ml).

Qu trnh nhn ging c bt u bng vic chuyn ging t ng nghim sang nhng

bnh tam gic cha sn mi trng nhn ging. Ngi ta thng nhn ging vo cc

bnh ln men dung tch t 1 lt cho n hng ngn lt. Nhit trong qu trnh nhn ging

duy tr khong 26 1oC v thi gian nhn ging mi cp khong 72 gi.

Ngi ta thng nhn ging penicillium trong mi trng c thnh phn sau:

Cao ng 2% Glucose2% lactose 0.5%

Nitrat amon 0.125% Sunfat magi0.025 sunfat natri 0.05%

Kaliphotphat monoboric 0.2% CaCO3 0.5%

Trong cng nghip sn xut khng sinh hin nay, thng l dng nhng chng bin

i gen. Cng ngh bin i gen to ra nhng chng siu tng hp khng sinh. Theo

Talaro (1993), t chng penicillin chrysogenum u tin ch c kh nng sinh tng hp

6mg/l, hin nay ngi ta c nhng chng bin i gen t chng gc c kh nng sinh

tng hp 85000ng/l penicillin.

3.2.2 Qu trnh ln men :

Qu trnh ln men trong mi trng lng bng phng php ln men chm sn xut

penicillin c vn hnh theo phng php ln men hai pha:

- Pha th nht nui thu sinh khi trong khong 2 3 ngy. Trong pha ny h si pht

trin rt mnh v cc cht dinh dng d ng ha s c t bo hp th rt mnh, tc

sinh sn ca nm xy ra rt nhanh, s to thnh penicillin mi bt u.

- Pha th hai ln mn thu sn phm. pha ny h si pht trin chm li, pH tng dn v

t n gi tr khong 7 7,5. Trong pha ny penicillin c to thnh vi mc cc

i.

25


26/37

Trong hu ht cc trng hp, khi ln men, ngi ta thay th phn ln (hoc hon ton)

ng lactose bng ng glucose. Lng glucose ny c th c b sung lin tc hay

bn lin tc nhng phi gim st cht ch nng glucose trong sut qu trnh vn hnh

pha duy tr nng glucose lun mc thch hp nhm va gi khi lng h si nnh, va m bo sinh tng hp nhiu penicillin.

Trong thc tin, trnh xy ra thiu ht nht thi glucose , ngi ta c th kt hp b

sung mt lng nh ng lactose (khi , nu cha b sung kp glucose th nm mc s

t iu chnh s dng ng lactose nn khng xy ra hin tng t phn h si).

Ngoi ngun nit trong nc chit ng, ngi ta thng s dng phi hp (NH4)2SO4

va cung cp thc n N v S, va s dng iu chnh pH trong qu trnh ln men (pH

dch len men ban u thng c iu chnh v khong pH = 6,5 6,8 bng dung dch

NaOH hoc H3PO4); nng NH4 + thng khng ch trong khong 0,3 0,4 kg/m3 dch

ln men. Cht ph bt thng s dng l cc loi du bo nh: m ln, du u tng,

du vng, du cmTin cht to nhnh phenylacetic trong ln men sn xut penicillin

G (hoc phenooxyacetic trong ln men sn xut penicillin V) c b sung lin tc (hoc

b sung gin on lm nhiu ln) trong sut thi gian pha ln men penicillin, duy tr

nng trong khong 0,1 1,0 kg/m3 dch (nu t qu nm mc s tng hp ng thi

nhiu penicillin khc, nu nhiu qu s gy c cho nm v tng cng thc y qu

trnh hydroxyl ha sn phm penicillin).

Nhit ln men pha u khng ch 30oC, sau sang pha sau gi 22 25oC. Tc

sc kh v khuy trn c iu chnh duy tr nng oxy ha tan trong dch trong

khong 30%. Trong iu kin trn thi gian ln men mi m thng ko di khong 144

180 gi. Kt thc qu trnh ln men ngi ta c gng lc sm dch ln men, lm lnh

ri chuyn sang cng on trch ly v tinh ch thu penicillin.

STT Thnh phn Mi trng 1 Mi trng 2 Mi trng 31 Cao ng 2.0-2.4 - 2.0-3.0

2Kh ht c du (lc, u

tng hng dng)- 2.0-2.4

3 Lactose 5.0 5.0 1.04 Glucose hoc hydol 1.0 1.0 1.05 Du thc vt 0.5-1.0 0.5-1.0 2.5-3.5

26


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6 Amon nitrat 0.4 0.4 0.47 Sulfat natri 0.05 0.05 0.058 Kali photphat 0.4 0.4 0.49 Magie sulfat 0.025 0.025 0.25

10 Natri hyposunfit 0.2 0.2 0.211 Canxi cacbonat 0.5-1.0 0.5-1.0 0.5-1.012 Tin cht 0.2-0.4 0.3-0.4 0.3-0.4

* c im v thit b ln men chm:

Qu trnh ln men sn xut penicillin ngy nay ch yu c tin hnh trong thit b ln

men chm ch to bng nhm thp chu n mn CT2 vi khuy trn kiu tuc-bin (gm

nhiu tng cnh khuy), kt hp b tr h vch dn dng trong thng). Cng sut khuy

trn tiu hao c thit k khong 3kW/m3/h. Khng kh nn v khun c cp vo

qua h ng phn phi kiu vng xoy hay kiu r qut c l lp t st di y (hay

pha di cnh tuc-bin). Bn trong thit b c lp t nhiu tng ng trao i nhit

kiu vng xon kt hp ng thi vi trao i nhit qua thnh thit b hai lp v, m

bo iu nhit hiu qu trong sut qu trnh ln men. Dung tch thit b ph bin trong

khong 150 300m3, h s y thng chn khong 80%V (ph thuc vo k thut

v thit b ph bt). Thit b nhn sn xut ging c dung tch khong 10%V thit b len

men, c thit k tng t v thng c ghp cng vi thit b ln men. Ton b thitb ln men sn xut, thit b nhn ging ln v h thng trang thit b ph tr c thit

k v lp t m bo c th v sinh v thao tc vn hnh theo ch v khun cao (tt

nht nn b tr sao cho c thap dng ch thanh trng ng thi cho ton b h thit

b ny). Cc thng s kim tra qu trnh ln men bao gm: pH mi trng, nng oxy

ha tan, nhit , hm lng sinh khi v tc bin thin lng sinh khi, s lng,

kch thc v cu trc pellet, nng cc cu t c cht, nng penicillin, thnh phn

kh thi v cc ch tiu kim tra v vi sinh vt. Vic gim st v iu chnh qu trnh ln

men c xy dng trn c s khai thc c hai kiu tng tc trc tuyn (online control)

v tng tc khng phn hi theo quy lut (offline control), ph thuc vo kh nng p

ng ca h thit b hin c. ng thi xu hng my tnh ha trong kim tra v gim st

qu trnh ln men ang dn chim u th trong sn xut cng nghip.

27


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Hnh 5. S h ln men dng cho sn xut penicillin.

* Hiu qu kinh t chung ca qu trnh ln men chm

Nng lc sinh tng hp v tch t penicillin trong dch ln men l kt qu ca mi tng

tc ng thi ca hng lot yu t cng ngh nh: hot tnh sinh tng hp ca chng,

cng ngh ln men p dng, cht lng nguyn liu, c tnh thit b v nng lc p ng

cc yu cu cng ngh ca thit b, ch gim st v iu chnh cc thng s cng

ngh, nng lc v k nng vn hnh ca cng nhn.... Vi ngun c cht chnh l glucoza

v ln men theo phng php chm, h s phn b nguyn liu d tnh khong 25%

glucoza c nm mc s dng tng hp h si, 65% ng c s dng duy tr

s sng st ca h si, cn li ch khong 10% c nm mc s dng tng hp

penicillin. H s s dng thc n nit v lu hunh tng hp penicillin tng ng l

20% v 80%. Nng penicillin G trong dch ln men nhng nm 80 - 90 ca th k XX

t khong 80.000 UI/ml (tng ng nng sut khong 40 - 50 kg penicillin G/ m 3 dch

ln men ).

3.2.3 X l dch ln men v tinh ch thu penicillin t nhin :

C ba phng php thu nhn v tinh ch penicillin t mi trng nui cy, l:

Trch ly bng dung mi hu c

Hp ph

Trao i ion

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Trong ba phng php trn th phng php trch ly bng dung mi hu c c s dng

nhiu hn c. phng php ny da trn nhng u im sau:

Mui ca penicillin rt d tan trong nc

Acid penicillic rt d tan trong dung mi hu c

Cng on x l dch ln men v tinh ch thu penicillin t nhin c tm tt trong s

hnh 6 , bao gm cc cng on chnh sau y:

Hnh 6. S tm tt cng on x l dch ln men thu penicillin t nhin

S khi qu trnh x l dch ln men :

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3.2.4. Lc dch ln men :

Mc ch : Penicillin l sn phm ln men ngoi bo. V vy, ngay sau khi kt thc qu

trnh ln men ngi ta thng tin hnh lc ngay gim tn hao do phn hu penicillin

v gim bt kh khn khi tinh ch, do cc tp cht to ra khi h si nm t phn.

Thit b lc: ph bin l thit b lc ht kiu bng ti hoc kiu thng quay. Thng

thng, ngi ta ch cn lc mt ln ri lm lnh dch ngay chuyn sang cng on

tip theo. Ch trong nhng trng hp rt c bit mi cn phi x l kt ta mt phn

protein v lc li dch ln th hai. Hin tng t phn h si nm thng ko theo hu

qu lm cho dch kh lc hn.

Thu hi sinh khi nm: Phn sinh khi nm c ra sch, sy kh v s dng ch

bin thc n gia sc.

3.2.5. Trch ly :

Penicillin thng c trch ly dng axt ra khi dch lc bng dung mi amylacetat

hoc butylacetat pH = 2,0 - 2,5, nhit 0 - 30C. Nhm hn ch lng penicillin b

phn hu, qu trnh trch ly c thc hin trong thi gian rt ngn trong thit b trch ly

ngc dng lin tc kiu ly tm nhiu tng cnh. ng thi, trong thi gian trch ly cn

gim st cht ch cc thng s cng ngh nh: nhit pH, v khun.... hn ch

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tn tht do phn hu penicillin. Dch ln men sau khi lc c bm trn ng thi vi

dung dch H2SO4 hoc H3PO4 long c b sung thm cht chng to nh v bm song

song cng vi dung mi trch ly vo trong thit b. T l dch lc: dung mi thng chn

trong khong 4 - 10V dch lc /1V dung mi. Trong mt s cng ngh, nhm ci thin

cht lng sn phm, ngi ta c th p dng phng php trch ly hai ln dung mi, viln u trch ly penicillin bng amylacetat hoc butylacetat; tip theo penicillin li c

trch ly ngc sang dung dch m pH = 7,2 - 7,5, thng l dung dch KOH long hoc

dung dch NaHCO3; sau penicillin li c trch ly sang dung mi ln th 2, vi lng

dung mi t hn.(hnh 7)

Hnh 7. S cng ngh trch ly 2 ln dung mi tinh ch penicillin

3.2.6 Ty mu :

ty mu v loi b mt s tp cht khc, ngi ta thng b sung trc tip cht hp

ph vo dung mi cha penicillin sau trch ly, s dng ph bin nht l than hot tnh.

Sau than hot tnh c tch v ra li bng s dng thit b lc ht bng ti hoc thit

b lc ht kiu thng quay. Phn than sau lc c a i chng thu hi dung mi v x

l hon nguyn, phc v cho cc m sau.

3.2.7 Kt tinh, lc, ra v sy thu penicillin t nhin:

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Vic kt tinh penicillin V hay penicillin G di dng mui c th c thchin rt n gin, bng cch b sung trc tip vo dung mi sau khi ty mu mtlng nh kali acetat (hay natri acetat) hoc ngi ta trch ly li sang dung dchKOH long (hay NaOH long), tin hnh c chn khng nhit thp, sau bsung BuOH penicillin t kt tinh. Cc thng s cng ngh c nh hng lnn hiu qa kt tinh l : nng penicillin, nng mui acetat, pH dung mihay pH dung dch c c, nhit kt tinh ... Sau khi kt tinh, tinh th penicillinc lc tch bng my lc ht thng quay. m bo tinh khit cao hn, cth tin hnh ha tan v kt tinh li penicillin. Khi sn phm t tinh schtheo yu cu, thng tinh khit khng di 99,5%, chng c lc tnh tinhth; tip theo ra v lm kh s b bng dung mi k nc nh izopropanol hay

butylalcohl; ht chn khng tch dung mi trn my lc bng ti ri sy bngkhng kh nng n dng sn phm bt mui penicillin. Sn phm ny, mt phnc s dng trc tip pha ch thuc khng sinh penicillin; cn li, phn lnc s dng lm nguyn liu phc v cho vic sn xut cc sn phm penicillin

v cephalosporin bn tng hp khc. Ngoi ra, sn xut ra cc sn phmpenicillin c tinh khit rt cao, ngi ta cn phi s dng phi hp thm mt sgii php cng ngh khc.

3.2.8 Sn xut penicillin bn tng hp t penicillin t nhin:

V nguyn tc, to ra mt penicillin mi trong ln men sinh tng hp c th

dng phng php thay i mch nhnh bng cch s dng cc tin cht khc

nhau. Tuy nhin bng con ng ln men trc tip hin nay ngi ta ch mi c

kh nng tin hnh vi penicillin G v V. Cn trin khai trong sn sut cng

nghip, ngi ta s dng penicillin G hay V ch to ra 6-APA l nguyn liu

ch yu trong qu trnh bn tng hp ra ccpenicillin mi.

Theo cu trc ha hc, cc penicillin tnhin u l nhng hp cht d vng

ca cc acid amin c tn gi l acid 6aminopenicillanic (6-APA). Vic tm ra

phn t 6-APA m ra mt hng mi sn xut penicilin bn tng hp bng

cch gn vo cc mch nhnh khc nhau vo phn t 6-APA bng con ng ha

hc hay sinh hc. C hai hng chnh l acyl ha nhm NH 2 v tr s 6 v esterha nhm COOH v tr s 3.

Enzyme penicillinamidase ng vai tr trong vic thy phn penicillin G hoc V

to thnh acid 6- aminopenicillanic (6-APA) v acid phenyl acetic. 6- APA l

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tin cht quan trng c s dng sn xut cc khng sinh h -lactam hot

ng cao.

6-APA c th sn xut bng phng php ln men cc chng penicillinum.

ng hc ca qu trnh ln men to 6-APA: ng ha hydratcarbon nhanh, pH

tng dn v trong giai on to 6- APA t 7.0 -7.4. Thi gian ln men ko di t

108- 120 gi. Tuy nhin qu trnh ny cho hiu sut thp v 6-APA c to thnh

ng thi vi cc penicillin t nhin , mc khc chi ph cho qu trnh tch chit

cao. V vy trong thc t ngi ta khng s dng phng php ny.

Hin nay s dng enzyme c nh l phng php ph bin sn xut 6-APA

trong cng nghip. S dng enzyme penicillin acylase ct mnh nhnh ca

penicillin G . Phng php ny c nhiu u im, iu kin phn ng d dng nn

mang li hiu qu kinh t cao. Hin nay mt s hng dc phm p dng bin

php ny, in hnh l hng dc phm Snam Progetti (Italia) vi sn lng

khong 40 tn/ nm.

Phng php enzyme c nh sn xut 6-APA cho hiu qu tng i cao (90-

95%) v c p dng rng ri nhiu nc, mang li hiu qu kinh t cao.

Hn 50% 6_APA trn th gii sn xut theo phng php enzyme c nh. Hinnay mt s nh my

sn xut

penicillin ch cho xut xng thnh

phm 6-APA.

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4. Cc thit b dng trong sn xut penicillin :

Thit b sy chn khng Thit b lc sinh hc

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Thit b lc than hot tnh Thit b sy

Khu vc ln men sn xut penicillin.

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Phn II Kt lun

Penicillin l lai khng sinh u tin c dng iu tr vim nhim ngi. Khm

ph v penicillin t mt pht hin tnh c nhng c ngha cc k to ln i vi nhnlai

Penicillin thng thng c sn xut bng phng php ln men chm a kh v c

chit xut pH thp t dch ln men s dng butyl axetat hoc kerosen. Do tnh

khng n nh ca penicillin pH thp, quy trnh chit xut cn thc hin nhit

thp, song c ti 10-15% sn phm b tht thot khi s dng quy trnh hin nay.

Ngoi nng sut tng i thp, cc phng php sn xut truyn thng c chi ph cao,

tiu th nhiu nng lng thu hi butyl axetat t cc cht l quy trnh i hi nhiucng on.

Penicillin c thi ra rt nhanh nhng ngi bnh nhn c chc nng thn bnh

thng, v c a vo c th qua con ng tnh mch nhng bnh nhn b nhim

trng nng. tc dng ngc quan trng v ph bin nht i vi Penicillin l d ng.

V kin thc cn hn hp nn c rt nhiu sai st mong c v cc bn cng ng gp

kin bi tiu lun hon chnh hn.

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Phn III:Ti liu tham kho

[1] Nguyn Vn Cch, Cng ngh ln men cc cht khng sinh, Nh xut bn Khoa hc

K thut, H Ni 2005[2] Nguyn Ln Dng, Nguyn nh Quyn, Phm Vn T, Vi sinh vt hc, Nh xutbn gio dc 1997[3] Trng Th Mimh Hnh, Gio n mn hc cng ngh dc phm , ti liu in t

[4] Nguyn c Lng, Vi sinh vt hc cng nghip, Nh xut bn i hc quc gia

TP.H Ch Minh

[5] Hnh nh t internet

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