Baj Moj Penicillin

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Trng i hc Tn c Thng

Khoa: Khoa hc ng dngNgnh cng ngh sinh hc

Seminar:

CNG NGH SN XUTPENICILLIN

SVTH: Tn Th Xun HuynNguyn Th Lan HngHong Hng NgcPhan Ngc Bo Nguyn


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Mc Lc

I. GII THIU CHUNG V CHT KHNG SINH:

1.1. Lch spht hin cht khng sinh

1.2. nh ngha khng sinh:

1.3. Cch tc dng:

1.4. n v khng sinh:

II. CHT KHNG SINH PENICILLIN:

2.1. Lch spht hin v sn xut penicillin

2.2. Cng thc cu to ca penicillin

2.3. Nhng vi sinh vt sn sinh Penicillin v c im dinh dng

ca chng.

2.4. Cscng ngh sinh tng hp penicillin tnm mc

III. QUY TRNH SN XUT PENICILLIN TVI SINH VT:3.1. c im chung:

3.2. Chun b ln men:

3.3. K thut ln men:

3.3.1. K thut ln men b mt:

3.3.2. K thut ln men chm:

3.4 Xl dch ln men v tinh ch thu penicillin tnhinIV. SN PHM:


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I. GII THIU CHUNG V CHT KHNG SINH:

1.1.Lch spht hin cht khng sinh:

Spht trin v vi sinh vt hc ni chung, v vi sinh vt cng nghip ni ring,

vi bc ngot lch s l pht minh v i v cht khng sinh ca Alexander

Fleming (1982) mra 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. Lin tip sau l nhng pht hin khc ca:

Babes (1885) nu ra nh ngha hot tnh khng khun ca mt chng l

c tnh tng hp c cc hp cht ho hc c hot tnh km hm cc chng i

khng.

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 tca loi trc khun

ny.

Gratia v ng nghip (1925) tch c tnm mc mt ch phm c th

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

Mc d vy, trong thc t 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.

1.2. nh ngha khng sinh:

Cht khng sinh c hiu l cc cht ho hc xc nh, khng c bn cht

enzym, c ngun gc sinh hc (trong ph bin nht l tvi sinh vt), vi c tnh

l ngay nng thp (hoc rt thp) c kh nng c ch mnh m hoc tiu

dit c cc vi sinh vt gy bnh m vn m bo an ton cho ngi hay ng vt

c iu tr.

1.3. Cch tc dng:

Cch 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


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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. Vtr tc dng chnh ca mt scht khng sinh

1.4. 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 sinhUI/ml (hay UI/g,International Unit.


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II. CHT KHNG SINH PENICILLIN

2.1. Lch s pht hin v sn xut

penicillin:

Penicillin c pht hin tnh cvo nm

1928 do Alexander Fleming, khi nhn thy mt

hp petri nui Staphylococcus b nhim nm mc

Penicillium notatum c xut hin hin tng vng

vi khun b tan xung quanh khun lc nm.

ng sdng ngay tn ging nmPenicillin t tn cho cht khng sinh

ny (1929).

Sau , M trin khai ln men thnh cngpenicillin theo phng php ln

men b mt (1931). Tuy nhin, cng trong khong thi gian mi n lc nhm

tch v tinh ch penicillin tdch ln men u tht bi do khng bo vc hot

tnh khng sinh ca ch phm tinh ch v do vn penicillin tm thi b lng

qun.

Nm 1938 Oxford, khi tm li cc ti liu khoa hc cng b, Ernst Boris

Chain quan tm n pht minh ca Fleming v ng ngh Howara Walter

Florey cho tip tc trin khai nghin cu ny.

Ngy 25/05/1940 penicillin c thnghim rt thnh cng trn chut.

Nm 1942, tuyn chn c chng cng nghip Penicillium chrysogenum

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

ny c xem l chng gc ca hu ht cc chng cng nghip ang sdng 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)....


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Hnh 2. Cc tc gi gii thng Nobel y hc nm 1945 v cng trnh

penicillin

Penicillin c xem l loi khng sinh ph rng, c ng dng rng ri trong

iu tr v c sn xut ra vi lng ln nht trong s cc cht khng sinh

c bit hin nay. Chng tc dng ln hu ht cc vi khun Gram dng v

thng c chnh iu tr trong cc trng hp vim nhim do lin cu khun,

t cu khun, th d nh vim mng no, vim tai - mi - hng, vim ph qun,

vim phi, lu cu, nhim trng mu...Thi gian u penicillin c ng dng iu

tr rt hiu qu. Tuy nhin, ch vi nm sau xut hin cc trng hp khng

thuc v hin tng ny ngy cng ph bin hn.

V vy 1959, Batchelor v ng nghip tch ra c axit 6-

aminopenicillanic. y l nguyn liu sn xut ra hng lot ch phm penicillin

bn tng hp khc nhau.

i vi Vit Nam, nm 1946, gio sng Vn Ng thnh cng trong vic

sn xut nc lc penicillin trong mi trng nc ng gp phn ng k vo vic

cu cha thng bnh binh v c Bc H thng Hun chng Lao ng

hng ba cho thnh tu k diu cha tng ai lm c ny.

2.2. Cng thc cu to ca penicillin:

Penicillin gm nhiu loi, chng c cu to gn ging nhau, bao gm mt vng

thiazolidine, mt vng -lactam, mt nhm amino c gn vi CO2 v mt mch bn(R).Tt c cc penicillin u l dn sut ca acid 6-aminopenicillanic. Sthay th R


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to nhiu acid amin khc nhau. hu ht cc penicillin u c phn phi di

dng mui natrii hoc mui kali.

Hnh 3. Cu to chung ca phn tpenicillin

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.

Hnh 2: Sn phm penicillin ln men tnhin nhP.chrysogenum

Cu trc khng gian ca Penicillin.


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2.3 Nhng vi sinh vt sn sinh Penicillin v c im dinh dng ca chng:

Nhng vi sinh vt sinh penicillin thuc cc ging nm mc penicillium v

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 scht tnhin

to thnh 10-15v/ml khng sinh.

Penicilillium chrysogenum trn mi trng Raistrik to thnh hai kiu khun

lc:

Kiu 1: khun lc trn trn, cc npnhn r nt. Khun ty kh sinhmc tt v c mu xanh, theo rakhun lc c ng vin rng 2-5 mm ca nhng

khun ty bc trng khng c bo t, cc khun ty ccht mu nu, cht mu khng

ha vo mi trng.

Kiu 2: Khun lc c nhng khun ty mu trng pht trin yu,khun ty ccht cng c mu nu. Khun lc kiu 1 cho hot lc cao, kiu 2 thng

xuyn cho hot tnh khng sinh thp. V vy cn phi tch nhng khun lc kiu 1

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

cao, gic c tnh ca ging.

Cc chng penicillium nui cy trn a petri


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cc chng Penicillium c nui cy trn a petri.

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

ny t cho cc nh vi sinh vt mt nhim v kh khn: to c kh nng sinhkhng sinh cao nht, gic n nh trong qu trnh nghin cu v sn xut.

Nhim v ny c mt ngha rt ln trong cng nghip, cc ging c bo vk,

trng thi ng kh c th ti 3 nm, t v trng l 2 nm. Ngy nay nhdi

truyn hc to ra c nhng ging n nh, t nht sau 6 th h khng lm gim

hot tnh khng sinh.

Penicillin thng 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 tging cbn v nhim

v ca cc nh vi sinh vt lcny l phi chn li nhng khun lc khe 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:

1. 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.

2. 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 .

3. Giai on III: To thnh nhng git cht bo to, khng cn khng

bo, t bo cht rt a kim.


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4. Giai on IV: Xut hin khng bo vi nhng ht d bt mu

trung tnh, nhng ht cht bo nh hn giai an III, tnh a kim gim.

5. Giai on V: Khun ty c hnh trng v c cha nhng khng bo,

gia c mt hoc mt vi ht ln. Cc ht cht bo bin mt. Tnh a kim tip tc

gim.

6. Giai on VI: Khun ty vn gic hnh dng hnh trng nhng

khng cn nhng ht bt mu trung tnh, cc khng bo bt mu da cam hocmu

hng ng u. Cc ht cht bo khng cn. Xut hin nhng t bo ring bit bt

u tphn.

Qu trnh ln men penicillin cng thuc vo loi ln men hai pha: pha sinh

trng (ng vi giai on I, II, III) v pha sinh penicillin ( cc giai on IV, V, VI ).

Ngun carbon trong ln men penicillin bng nm penicillium chrysogenum

c th l glucuza, sacaroza, lactoza, tinh bt, dextrin, cc axit hu c(lactic, axetic,

formic), cc axit aminng lactoza cho hiu xut penicillin cao nht v thng

c dng trong cng nghip. nm thng sdng lactoza chm v vy, trong thc

t lactoza c dng phi hp cng ng khc (glucoza, sacaroza) trong mi

trng dinh dng.

Trong pha ln men thnht ging pht trin mnh, sdng glucoza v axit

lactic ca cao ng. Sau lactoza mi oc s dng ( ch yu trong pha to

penicillin). Khi trong mi trng cn lactoza v khng b sung cc cht dinh dng,

h si nm bt u tphn, nu tip tc ln men nng pecicillin s gim, trong

thc t cn kt thc trc thi im ny.

Ngun nit: c th l nhng hp cht hu c(axit amin, pepton, protein) v

v c (amoniac, cc mui amon v nitrat). Amoniac c nm penicilliumchrysogenum ng ha nhanh hn c. trong qu trnh nui cy N-NH3c to

thnh tcao ng do phn ng khamin cc hp cht nit. Nm mc sdng N-

NH3 trc tin v nng ca cht ny trong thi gian u tng ln, v tc sinh

trng, pht trin ca nm mc v tip tc gim cho n khi h si ca mc t

phn. Tc sdng amoniac ph thuc ngun carbon trong mi trng. Trong

trng hp ngun carbon l glucoza, sacaroza hoc ngun carbon d tiu ha khc,


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amoniac sdng nhanh hn khi mi trng coa lactoza. Nitrat c nm mc ng

ha khi trong mi trng khng co ngun nithu c.

Lu hynh c ngha c bit quan trng trong qu trnh sinh trng v

sinh tng hp ca nm mc. ngun lu hunh thng dng l mui sunfat ca kali,

natri v amon. Cc cht ny tham gia vo tng hp metionin, sixtin, biotin,

tiamin hoc trng thi lin kt yu l tt hn c. Nhiu cng trnh nghin cu cho

bit, khi trong mi trng c mt ng thi L-sixtin v sufat th lu hunh ca axit

amin ny di vo phn tpenicillin hn lu hynh ca cc gc sufat. Song, dng

axit amin trong sn xut khng kinh t cho nn ngi ta thng dng thiosufat

natri (Na2S2O3). Lu hunh ca cht ny rt d di ng. Trong mi trng dinhdng c thiosufat cng vi cao ng hiu sut penicillin c th tng hai ln.

C cht bin i cc hp cht lu hunh t dng oxy ha sang dng kh

theo s ca Arnstein (1954) nhsau:

Sufat Sufit Thiosufat Sixtin

Trong t bo sixtin d bin thnh sixtein v ngc li. sixtein c mt ngha

ln nhmt tc nhn khnhnhm sunfuhydrin (-SH)

pH mi trng thch hp cho penicillium chrysogenum pht trin nm trong

khang 6-6.5. mi trng kim hac axit hn u lm cho mc pht trin chm.

trong qu trnh ln men pH mi trng thay i ty thuc vo tc sdng cc

hp cht cacbon v N-NH3.

2.4. Cscng ngh sinh tng hp penicillin t

nm mc:

2.4.1. Lch stuyn chn chng cng nghip

P. chrysogenum:

Vo nhng nm u, vic nghin cu sn xut

penicillin thng sdng cc chng c hot lc cao

thuc loi P. notatum v P. baculatum. Nhng tkhi

trng i hc Wisconsin (M) phn lp c

chngP.chrysogenum c hot tnh cao hn th chng

ny dn dn thay th v t khong sau nhng


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

sdng 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 cbn m t trong mc 1.3.1, trong gii php

k thut c p dng hiu qu thu nhn bin chng "siu tng hp"

penicillin li chnh l cc k thut gy t bin thng nh: xl tia Rn - ghen, x

l tia cc tm v to t bin bng ho cht, th d nhMetylbis - amin

(metyl -2--clo- etylamin), N-mustar (tris - -clo- etylamin), Sarcrolyzin, HNO2,Dimetylsulfat, 1,2,3,4 -diepoxybutan.

2.3\4.2. Cch sinh tng hp penicillin nm mc P. chrysogenum :

Theo quan im ph bin hin nay, qu trnh sinh tng hp penicillin nm

mc P. chrysogenum c th tm tt nh sau: t ba tin cht ban u l -aminoadipic, cystein v valin s ngng t li thnh tripeptit -(- aminoadipyl) -cysteinyl - valin ; tip theo l qu trnh khp mch to vng -lactam v vngthiazolidin to thnh izopenicillin-N; ri trao i nhm -aminoadipyl viphenylacetic (hay phenooxyacetic) to thnh sn phm penicillin G (hay penicillin

V, xem s tng hp penicillin G trong hnh.

Hnh 3. S

cch sinh tng hp penicillin taxit L-- aminoadipic, L-cystein v L-valin


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Tuy nhin, cng c th n c gii phng ra v tch t trong mi trng (v

trong qu trnh ln men sn xut penicillin V bao gi cng pht hin thy trong

dch ln men lng ln - aminoadipic dng vng). Nhvy, qu trnh sinh tnghp penicillin, ph thuc vo iu kin ln men c th nht nh, c th xy ra theo

su ng hng khc nhau. Do , hiu sut chuyn ho ccht - sn phm cng

bin i v ph thuc vo ng hng sinh tng hp tng ng. Theo l thuyt th

hiu sut ln men s trong khong 683 - 1544 UI penicillin/g glucoza; song, trong

thc t, vi nhng chng c hot tnh sinh tng hp cao nht cng mi ch t

khong 200 UI/g glucoza.

2.4.3. Tc ng ca cc thng s cng ngh n qu trnh sinh tng hppenicillin.

2.4.3.1. Spht trin h si v c im hnh thi h si nm:

Spht trin hsi nm trong qu trnh ln men bao gm:

-Stng trng vkch thc hsi(tng di si, sln ln v kch thc,

mc phn nhnh ca h si ... )

- Sbin thin vslng khm si nm trong mi trng: Thng thng, s

pht trin ny c nh gi qua hai ch tiu l: hm lng sinh khi v tc bin

thin hm lng sinh khi trong mi trng. Hai ch tiu ny c th xc nh bng

nhiu phng php khc nhau nh: hm lng sinh khi (Sinh khi ti hoc sinh

khi kh), mt quang dch ln men, trlc 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 nh

qua hm lng sinh khi.

Tc pht trin h si nm ph thuc hng lot cc yu t khc nhau trong

qu trnh ln men v stch t penicillin thng xy ra mnh m khi h si pht

trin t trng thi cn bng. Trng thi ny c th xc lp c khi ch cung cp

va v lin tc lng thc n ti thiu cho nm mc. Thiu thc n, h si nm

s tphn, cn nu cung cp qu nhu cu trn, h si s pht trin, nhng khng

tch t mnh penicillin m tch t nhiu axit gluconic v axit malic.


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- 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 gicng c xu hng

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 chc vi cnh khuy v cc chuyn ng dng

xoy trong mi trng, mt mt st gy h si nm xy ra nhiu hn v h si

nm bao gicng c xu hng vn cun li thnh cu trc bi si cun xon, c

gi l pellet.

Pellet x p (fluffy loose pellets) l dng pellet c phn bn trong h si cunthnh 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 tphn tothnh 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 cvi s lng, kchthc 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 dso 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 iu kin cn i vi nhu cu tit kim mc tiu tn

nng lng do khuy trn.

2.4.3.2. c tnh nhit ng ca dch ln men:

Trong cc thit b ln men dung tch ln c sc kh v khut trn, thc t

khng th xc lp c sng 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


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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 ca thit b c th xut hin vng xoy cc b hay cc dng chy

thcp 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 dtrn mc yu cu.

2.4.3.3. Thnh phn mi trng ln men:

Mi trng cs ln men penicillin, vo thi ku trong nhng nm 40 -

50, l mi trng lactoza - nc chit ng, vi thnh phn chnh nu trong bng 2.1.

Ngun ccht chnh: l lactoza c thc 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 ccht nu trn, hiu qu cao

hn c vn l glucoza.

Ngoi ra, khi sdng du thc vt lm cht ph bt phi xt n hiu ng

nm mc sdng mt phn du thc vt lm ngun cung cp thc n cacbon,

tnh ton iu chnh nng glucoza trong mi trng ln men (v c scn

trqu trnh chuyn khi do nh hng ca du ph bt).

Ngun cung cp thc n nit: c th sdng l bt u tng, bt ht

bng, cc loi du cm. Nhu cu v thc n nitcng c thc p ng bng

cch cung cp lin tc (NH4)2SO4, nhng duy tr nng thp, khong 250 -

340g/l (nu dtha hiu qu sinh tng hp penicillin s gim, nu thiu s xy

ra hin tng tphn h si) .

Hm lng cc cht khong b sung: c tnh ton, ph thuc vo lng

dch chit ng sdng

pH mi trng c iu chnh trc khi thanh trng, sau trong sut

qu trnh ln men c gim st cht ch v iu chnh theo yu cu cng ngh.


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Nng tin cht to nhnh:Trong qu trnh sinh tng hp penicillin, vic

kt gn mch nhnh ca phn tpenicillin khng mang tnh c hiu cht ch.

Nhvy, nu duy tr nng tin cht to nhnh cn thit phenylacetat (hoc

phenooxyacetat) s cho php thu nhn ch yu mt loi penicillin G trong dch

ln men (hoc penicillin V). Theo l thuyt, nhu cu v phenylaceta l 0,47g/gam

penicillin G (hoc phenooxyacetat l 0,50g/gam penicillin V ). Cn ch c hai

cu ttrn 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 ln men ca chng sn xut.

2.4.3.4. iu kin tin hnh ln men:

Nhit l thng s c nh hng ln n spht 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


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ging sn xut ngi ta thng nhn di nhit 300C; sang giai on ln men

thng p dng mt trong hai ch nhit l :

Ln men mt di nhit : Thng duy tr nhit trong sut qu trnhln men di nhit 25 - 270C.

Ln men hai ch nhit : Giai on ln men bt u tin hnh 300Ccho n khi h si pht trin t yu cu v hm lng sinh khi th iu chnh

nhit sang ch ln men penicillin di nhit 22 - 250C (c cng nghiu

chnh xung 22 - 230C, ginhit ny tip hai ngy ri chuyn sang ln men tip

250C cho n khi kt thc qu trnh ln men).

pH mi trngthun li cho spht trin h si v cho qu trnh sinh tng

hp penicillin thng dao ng trong khong pH = 6,2 - 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 chgi 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 tnm mc; tuy nhin nu nng CO2 qu cao s lm cn tr

qu trnh hp thu v chuyn ho c cht ca chng, ngha lm lm cn tr qu

trnh sinh tng hp penicillin.

2.4.3.5. Stch 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 nnglc 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.


18/36


III. QUY TRNH SN XUT PENICILLIN TVI SINH VT:

3.1. c im chung:

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

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 nhsau (xem s hnh 2.8)

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

Xl dch ln men tinh ch thu bn thnh phm penicillin tnhin.

Sn xut cc penicillin bn tng hp (tnguyn liu penicillin tnhin)

Pha ch cc loi thuc khng sinh penicillin thng mi

Ging penicilliumchrysogenum

Nhn ging nh

Nhn ging sn xut

Ln men sn xut

Lc

Trch ly

Hp ph bngthan hot tnh

Sinhkhi

Dung mi


19/36


S sn xut penicillin (theo Gist-Brocades Copr (H Lan)).

3.2. 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 th.

Khi mi pht hin trong mi trng c, chng to ra hai dng khun ty: khun ty kh

sinh v khun ty dinh dng mu trng. Sau khi nui cy mt ngy, khun ty bt u

chuyn sang mu xanh xm v nh bo tbt u xut hin. Thi gian ny xut hin

mt t bo ttrn ttin bo tnm trong cc nh bo t. Cc bo tln lt c to

thnh theo thi gian nui cy v cui cng th mu ca nm penicillium sm hn.

Lc

Kt tinh penicillin

Than

Lc tinh th

Ra tinh th

Xy kh

Sn xut penicillinbn tng hp

Pha ch penicillinthng mi

Hp ph bng

than hot tnh


20/36


Ging cng nghipP.chrysogenumc bo qun lu di dng ng kh, bo

qun siu llnh hoc bo qun trong nitlng. Ging tmi trng bo qun c

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

huyn ph bo tthu thp 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 m bo ng thi im cho 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.

- Chun bmi 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

thng cha 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 thanh

trng ng thi dch ln men trong cng mt thit b. Tt c cc cu tb sung vo


21/36


mi trng ln men u phi c x l khkhun trc v sau b sung theo

ch vn hnh v khun.

Thit b ln men: Phi c v khun trc khi a vo sdng. Thngthanh 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 trkhc.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 khkhun sb bng nn on nhit, sau qua

mng lc v khun hay mng siu lc .

- Chun bmi trng nhn ging

Chun b mi trng nhn ging: lm mi trng nhn ging ngi tacng chun b nhmi trng ln men nhng chng khng cha lactose (nu c ch

cha mt lng rt nh), mt s khong cht v tin khong cht. Mt khc 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 shnh thnh

v pht trin thun li ca pellet.

Sau khi lm m mi trng n m nht nh, ngi ta s phn phi vo

chng vo cc dng c thy tinh (chai thy tinh hay cc bnh tam gic) vi khi

lng bng 1/5 hay 1/6 dung tch ca dng c, y nt bng v em thanh trng

121oC (0,5 at) trong 30 pht.

3.3. K thut ln men:

3.3.1. K thut ln men b mt:

p dng t lu, hin nay hu nhkhng cn c trin khai trong sn xut

ln na. Gm 2 phng php:

* Ln men trn nguyn liu rn (cm m, cm ng c b sung ng lactose)

* Ln men trn b mt mi trng lng tnh (ph bin sdng mi trng c

bn lactose - nc chit ng)..


22/36


S CNG NGH LN MEN NI PENICILLIN

Qu trnh nhn gingQu trnh nhn ging bt u tging 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 ng dng sn xut v mt ng dng bo qun.

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

chun b 50 g mi trng nhphn trnh by trn. 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 tha trn vi nc. Bng bin php v trng (thc hin trong cc t nui

cy v trng) chuyn ton b vo bnh tam gic trn lc cho tht u v chuyn

chng sang tm 30 37oC. Nui trong iu kin ny chi n khi bo tnm xut

hin v pht trin u khp mi trng.

Ta gi qu trnh thc hin nhtrn l qu trnh nhn ging cp 1. Cln lt

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

Ging penicilliumchrysogenum

Nhn ging

Ln men

Tch v tinhch

Sn phmpenicillin

Nguyn liu

Chun b mitrng ln

men

Chun b mitrng nhnging


23/36


Mi mt cp nhn ging tcp ny sang cp khc, khi lng mi trng tng

t10 15 ln. trong trng hp qu mt k ngi ta nui trn nhng khay.

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

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

Theo Talaro (0993), tchng penicillin chrysogenum u tin ch c kh nng tng

hp 6mg/ml, hin nay ngi ta c nhng chng bin i gen tchng gc c kh

nng sinh tng hp 85000mg/ml penicillin.

Qu trnh ln meni vi phng php ln men trn nguyn liu rn (cm m, cm ng c b

sung ng lactose), khi mi trng c khtrng v lm ngui n 30oC, tin

hnh trn ging vo vi t l t5 10%. Cc khay c xp chng ln nhau trn

nhng gi c mt khong cch nht nh thong kh v thong nhit. Qu

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

Nm penicillinum trong qa 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.

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

KH2PO4 0.5g

C6H5CH2COOH 0.2g

Lactose 30g

MgSO4 0.25g

ZnSO4 0.044g

NaNO3 3g

Nc 1000g

u im ca phng php ny l ng lactose c nm mc ng ha

chm nn khng xy ra hin tng dtha ng trong t bo, cn dch nc chit

ng cung cp cho nm mc ngun thc n nit, cc cht khong v cc cht sinh

trng, trong phenylalanin khi b thy phn s to thnh phenylacetic cung cp

tin cht to mch nhnh cho phn tpenicillin.

Khi ln men trong mi trng lng, p dng cng ngh b sung lin tc

phenylacetic vo mi trng ln men, hm lng b sung ph thuc pH mi trng

thng l 0,2-0,8 kg phenylacetic/m3 dch ln men. Dung dch ln men sau khi c

khtrng sc phn phi vo cc khay c kch thc ging cc khay nui cy b


24/36


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 6 7 ngy nhit ln

men l 24 28oC. Tin hnh ln men trong iu kin mi trng lng ny, lng

penicillin G c tng hp tng r rt cn hm lng cc penicillin khc cng gim

i. hn ch qu trnh oxy ha tin cht, thng phi b sung vo mi trng mt

lng nh axit axetic. Trong k thut ln men lng gin on khng iu chnh pH

mi trng thng tng nh, sau tng i n nh v vo cui qu trnh ln

men thng trong khong pH = 6,8 7.4. Khi s dng c cht chnh l lactose,

ngi ta xc nh c penicillin chc tng hp v tch t mnh m trongmi trng khi nm mc sdng ng ny v khi lactose c du hiu cn kit

th si nm cng bt u tphn. V vy ngi ta thng kt thc qu trnh ln

men vo thi im sp ht ng lactose.

3.3.2. K thut ln men chm:

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

xut penicillin cng nghip hin nay thay th dn k thut ln men ni doc u m

hn v thng c vn hnh theo phng php ln men bn lin tc, gm phngn ln men gin on theo m c b sung lin tc (hay bn lin tc) mt hay mt vi

cu tkt hp vi phng n tun hon li mt phn h si ca m ln men trc

(hoc khng).

Trong phng php ln men chm th mi trng c sdng l mi trng

lng, ngi ta dng cao ng, glucose, hydrol,lactose, cc mui amon, thiosufat,

photphat kali hoc Natri, cc mui sufat magie, natri, ng

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 tng nghim

sang nhng bnh tam gic cha sn mi trng nhn ging. Ngi ta thng

nhn ging vo cc bnh ln men dung tch t1 lt cho n hng ngn lt. Nhit


25/36


trong qu trnh nhn ging duy tr khong 26 1oC v thi gian nhn ging mi

cp khong 72 gi.

Ngi ta nhn ging penicillin trong mi reng vi ccthnh phn sau:

Cao ng 2%

Nitrat amon 0.125%

Kaliphotphat monoboric 0.2%

Glucose 2%

Sunfat magie 0.025%

Lactose 0.5%

Sunfat natri 0.05%

CaCO3 0.5%

Qu trnh ln menQu 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 thnht nui thu sinh khi trong khong 2 3 ngy. Trong pha ny h

si pht trin rt mnh v cc cht dinh dng dng ha sc t bo hp th

rt mnh, tc sinh sn ca nm xy ra rt nhanh, sto thnh penicillin mi bt

u.

- Pha thhai ln men 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.

Ging nm mc penicillium chrysogenum l loi hiu kh bt buc, tuy nhin

trong qu trnh tng hp penicillin xy ra trong iu kin hiu kh mnh nn trong

sut qu trnh ln men vic thi kh l iu cn thit

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 thc 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 gikhi lng h si n nh, 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 nhng lactose (khi , nu cha b sung kp glucose

th nm mc s tiu chnh sdng ng lactose nn khng xy ra hin tng

tphn h si). Ngoi ngun nit trong nc chit ng, ngi ta thng sdng

phi hp (NH4)2SO4 va cung cp thc n N v S, va sdng iu chnh pH

trong qu trnh ln men (pH dch len men ban u thng c iu chnh v


26/36


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 sdng

l cc loi du bo nh: mln, 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 ch30o

C, sau sang pha sau gi22 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.

* c im v thit b ln men chm:

Qu trnh ln men sn xut penicillin ngy nay ch yu c tin hnh trongthit 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 hng 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 tv thng c ghp cng vi thit b ln men. Ton b thit b ln men

sn xut, thit b nhn ging ln v h thng trang thit b ph trc 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 th p dng ch thanh trng ng thi cho ton b h


27/36


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 tccht, 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 skhai 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.

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

* Hiu qu kinh t chung ca qu trnh ln men chmNng 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 iuchnh cc thng s cng ngh, nng lc v k nng vn hnh ca cng nhn.... Vi

ngun ccht chnh l glucoza v ln men theo phng php chm, h s phn b

nguyn liu dtnh khong 25% glucoza c nm mc sdng tng hp h si,

65% ng c sdng duy tr ssng st ca h si, cn li ch khong 10%

c nm mc sdng tng hp penicillin. H s sdng thc n nitv lu

hunh tng hp penicillin tng ng l 20% v 80%. Nng penicillin G trong


28/36


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/ m3 dch ln men ).

3.4 Xl dch ln men v tinh ch thu penicillin tnhin:

C ba phng php thu nhn v tinh ch penicillin tmi trng nui cy,

l:

Trch ly bng dung mi hu c Hp ph Trao i ion

Trong ba phng php trn th phng php trch ly bng dung mi hu cc sdng 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 tnhin c tm

tt, bao gm cc cng on chnh sau y:


29/36


Dch lnmen

Lc than

Lc si nm

Trch ly

Trch ly

pH 2,0-2,5

Kt tinh

Lc tinh th

Hp ph bngthan hot tnh

Sinh khinm

Thanhot tnh

Dung mi,

ph gia, axit.

Kali acetat hocNatri acetat

Ra tinh th

Tinh ch thuhi penicillin

Dung mi

Dung dch mpH= 7,2-7,5.


30/36


3.4.1. 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 tphn.

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 thhai. Hin tng tphn 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 sdng

ch bin thc n gia sc.

3.4.2. 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 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, vi ln u trch ly penicillinbng 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 th2, vi lng dung

mi t hn.


31/36


3.4.3. 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, sdng ph bin nht l

than hot tnh. Sau than hot tnh c tch v ra li bng sdng 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 xl hon nguyn, phc v cho cc m sau.

3.4.4. Kt tinh, lc, ra v sy thu penicillin tnhin:

Vic kt tinh penicillin V hay penicillin G di dng mui c thc thc

hin rt n gin, bng cch b sung trc tip vo dung mi sau khi ty mu mt

lng nh kali acetat (hay natri acetat) hoc ngi ta trch ly li sang dung dch

KOH long (hay NaOH long), tin hnh c chn khng nhit thp, sau b

sung BuOH penicillin tkt tinh. Cc thng s cng ngh c nh hng ln n

hiu qa kt tinh l : nng penicillin, nng mui acetat, pH dung mi hay pH

dung dch c c, nhit kt tinh ... Sau khi kt tinh, tinh th penicillin c lc

tch bng my lc ht thng quay. m bo tinh khit cao hn, c th tin

hnh ha tan v kt tinh li penicillin. Khi sn phm t tinh sch theo yu

cu, thng tinh khit khng di 99,5%, chng c lc tnh tinh th; tip theora v lm kh sb bng dung mi k nc nhizopropanol hay butylalcohl; ht

chn khng tch dung mi trn my lc bng ti ri sy bng khng kh nng n

dng sn phm bt mui penicillin. Sn phm ny, mt phn c sdng trc tip

pha ch thuc khng sinh penicillin; cn li, phn ln c sdng 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 phm penicillin c tinh khit rt cao,ngi ta cn phi sdng phi hp thm mt s gii php cng ngh khc nh

phng php phn tn tnh in. nguyn tc ca phng php ny l sdng mt

hiu in th cao (ln n 25 kV) to nhng vi git ca dung dch cha penicillin.

3.5 Sn xut axit 6- aminopenicillanic v sn xut penicillin bn tng hp

Axit 6- aminopenicillanic tuy khng c hot tnh khng khun, nhng c th s

dng lm nguyn liu tng hp ra nhiu loi penicillin khc nhau v c


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cephalosporin. sn xut axit 6- aminopenicillanic, con ng hiu qu hn c

hin nay l ln men sn xut penicillin G (hoc penicillin V); sau p dng

phng php ha hc hay sdng enzym acylaza phn ct mch nhnh bn xem

s hnh 2.11.

Hnh 2.11. S tng hp axit 6- aminopenicillanic tpenicillin G

Phng php ha hc c hiu sut chuyn ha cao, ti 90-95%, v tc phn

ng nhanh, song li tiu hao nhiu nng lng, nhiu dung mi v cha ng nguy

c nhim mi trng cao. Trong khi , phng php enzym, tuy hiu qu chuyn

ha thp hn nhng iu kin phn ng m du v mang li hiu qu kinh t cao

hn nn c trin khai ph bin trong thc tin sn xut cng nghip. ng thi,

cng nhu th trn, phng php enzym c hon thin lin tc v ngy nay

t c u th vt tri so vi phng php ha hc.

3.6 Tnh hnh sn xut penicillin trn th gii:

Thuc khng sinh chim khong 30% th phn dc phm th gii. Cc tp

on dc phm ca M, Anh, c, Php, Nht Bn, Italia, H Lan, Trung Quc,

n , i Loan, Hn Quc, o chim cc v tr hng u trong lnh vc ny. V

d: cng ty DSM c tr sthnh ph Delf (H Lan) hng nm sn xut 15.000 tn


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penicillin, chim 30% sn lng loi thuc ny sn xut trn ton th gii. Ngoi ra

DSM cn sn xut cc tin cht 7 ADCA (9.000 tn), 6 APA (hng ngn tn), cng

vi ampicillin, amocixillin, cephalexin, cefaroxin, cefradin... Tp on dc phm

HoaBc Thch Gia Trang ca Trung Quc sn xut mi nm 4.000 tn penicillin.

Tp on dc phm Thnh mi nm sn xut 3.000 tn penicillin, 500 tn 6

APA v 200 tn 7 ADCA.

THIT B SDNG TRONG SN XUT PENICILLIN

Thit b sy chn khng

Thit bi lc sinh hc


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thit b lc than hot tnh

thit bi sy


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Thit b sy v mt s sn phm ca n

IV. SN PHM:1.Penicillin nhm G:

y l nhm penicillin cin, tm thy u tin. a s trch tnm vmt s bn tng hp. cc penicillin Gc tac dng vi khun gram dng v cu

khun gram m. Sau60 nm sdng penicillin G phn no mt hiu lc do tora nhiu chng ln thuc.a s penicillin G b acid ca dch v ph hy nn ch dng tim m

khng ung.Thuc chn lc tr ph cu, lin cu, t cu khng tic penicillinaz,

clostridium, neisseria, khun k khming , xon cu.G.1_ Benzylpenicillin = penicillin G

Bt mu trng, mi vc bit, d tan trong nc.Dung dch thuc d b thy phn. Ch pha khi dng. Dc in

qui nh dung dch penicillin G chc tn tr3 ngy trong t lnh. Nhit cngcao cng d b hng

G.2_ Penicillin G-procainy l benzylpenicillin-procain c tc dng chm.c ch: epocilin, allocillin, duracillin,hydracillin, luetoben, novocillin, solucillinBt trng , khng ht m, t tan trong nc

G.3_BipenicillinV penicillin-procain khuch tn chm ngi ta kt hp penicillin G vi penicillinprocain c tc dng va tc thi va lu bn

G.4_Benzathin-penicillinBenzathin-penicillin = benzathin-benzylpenecillinc ch: extencillin, benapen, leomyben, benadin, tadocillin.Kt hp 2 phn tpenicillin G vi 1 phn tdibenzyl-etylendiamin.

G.5_Benethamin-penicillinBt kt tinh, t tan trong ncTc dng chm, trung bnh

G.6_Clemizol-penicillinc ch: lacgobenyl, neocuroClemizol l cht khng-histamin, nhng mui clemizol ca acid benzylpenicillinic ldng thuc c tc dng chm. shin din ca clemizol lm gim bt dng v tnguy cgy tai bin.

G.7_Penethacillinc ch neobenyl, leocillin, extoben, bronchocillin.Bt trng t tan trong nc


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G.8_Penicilln V = Phenomycillin = phenoxymethyl- penicillinc ch: oracillin, alphacillin, berocillin, calciopen K, iciben, isocillin, oracil,oragen, penigen, sumagen, pantogen, calcipen V, penavlon V

G.9_Phenethycillin = phenoxyethyl-penicillinc ch: proxyl, chemipen, maxipen, syncillin

G.10_Propicillin = phenoxypropylc ch: baycillin.

G.11_ClomethocillinTc dng lu di hn cc penicillin ung khcPenicillin nhm M 9 ( ly meticillin lm cn bn)Nm 1957, J.C Sheehan tng hp c penicillin. Nm 1929, Doyle v Robinson a vo sn xut cng nghip. bng cch thay i dy ngang R, ngi ta tmc nhiu cht mi c hot ph rng, t gy trngi khi dng, t to chng khng iu ng ch nht l tng hp c cc cht khng penicillinaz do t

cu tit ra v nhng penicillin ung tt hn penicillin V.Penicillin nhm M c 2 cht chnh l meticillin v clozalin. Chng khng b thyphn bi penicillinaz. Do cc cht ny c dng tr cc bnh t cu.Tuy nhin khong 20-25% chng t cu vn khng vi nhm penicillin M, ldo khng nhim th. Do cc penicillin ny vn cha c gi tr tuyt i vi tcu.Hiu lc khng khun ch bng 1/10 penicillin G vi cc khun nhy cm vipenicillinOxacillin, cloxacilin, diclozacilin t c tnh hn meticillin. Meticillin ch dng cccht khc ung c.Penicillin nhm A ( ly ampicillin lm cn bn):

y l penicillin bn tng hp, hot ph rng hn cc penicillin trc kia: dit ccc khun gram dng v m, cu khun gram m, nhng li tr khng hi hu tcuNgi ta chia thnh 3 nhm nh

Aminopenicillin: amicillin v cht dnCarboxypenicillin: carbonycillin v dn xut.Ureidopenicillin v imidinopenicillin

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