Post on 06-Jul-2018
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The Industrial Production of Penicillin
Pranjal Singhal
2012B1A1699G
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Overvie
● What is penicillin?● History
● General structure of penicillins
● Fermenters
●
Important aspects of fermentors● Specific conditions for
production
● Media consideration &
formulation
● Production ofpenicillin
● Increase in yield
● References
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!hat is Penicillin"●
First true naturally-occurring antiiotic e!er disco!ered" a greatmedical rea#through$
% Group of antiiotics produced y the Penicillium fungi$
It is a group of closely related compounds not a single
compound$Examples: 'mo(icillin ampicillin pheno(ymethylpenicillin$
% 'round )* drugs that are penicillins$
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#istor$% &iscover$ ' Production
● +,." Scottish iologist 'le(ander
Fleming disco!ered that the Staphylococcus
culture hehad mista#enly left gro/ing in open /as hadcontaminated /ith a mold /hich
destroyed the acteria$
● 'fter isolating a sample and testing it he
found that it elonged to the Penicillium
family$
0ater the mold /as classified as Penicillium
notanum.
● 't first it /as difficult to con!ince people
aout its potential uses$
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#istor$% &iscover$ ' Production
● 1ut later 2+,3,4 using Fleming5s /or# t/o medical
researchers Ho/ard Florey and 6rnst 7hain
managed to purify penicillin in a po/dered form$
% +,8+" 9hey successfully treated a human$
% +,83" 9hey produced penicillin on a large scale$
9his helped immensely to treat casualties during the
WWII that had acterial infections due to their
/ounds$
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General Structure of Penicillins
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(edia )onsideration
● 9he aim of the media is to"
of cell
○ pro!ide all the elements re:uired for the
synthesis materials and the formation of the desired
product$○ pro!ide fa!ourale en!ironment for the culture in :uestion$
○ e cost effecti!e$
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(edia *or+ulation
% pH ;$)
● 9emperature *-8 itrogen" corn steep li:uor .$)
● Glucose +● .* ethanol
● phenylacetic acid
● Proenecid
● 0actose +
● 7alcium 7aronate +
● Sodium hydrogen phosphate *$8
● 'ntifoaming agent" !egetale oil
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Si+,lified *lo )hart
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Source" Pfi@er
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★ (ediu+ for
,enicillin+$ 9he Penicillium chrysogenum usually contain its caron source
/hich is found in corn steep li:uor and glucose$
$ ' medium of corn steep li:uor and glucose are added to the
fermenter$ Medium also consists of salts such as MgS=8
A 3
P=8
and
sodium nitrates$ 9hey pro!ide the essential ions re:uired for the
fungus metaolic acti!ity$
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★ #eat
sterili-ation3$ Medium is sterili@ed at high heat and high pressure usually through a
holding tue or sterili@ed together /ith the fermenter$
8$ 9he pressuri@ed steam is used and the medium is heated to ++
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★
*er+entation)$It is done in a fed-atch mode as glucose must not e added in highamounts at the eginning of gro/th 2/hich /ill result in lo/ yield of
penicillin production as e(cessi!e glucose inhiit penicillin production4$
;$9he fermentation conditions for the Penicillium mold usually
re:uires temperatures at *-8
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★
*er+entation.$ It is necessary to mi( the culture e!enly throughout the culturemedium$ Fungal cells are ale to handle rotation speed of around **
rpm$
Fermentors
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1. Reactor Size
2. Reactor Configuration3. o!e of operation
". Con!itions insi!e t#e reactor
Economic requirements:%$ 6asy to operate aseptically$
%$ Reasonaly fle(ile regarding process re:uirements$
%$ 0o/ po/er consumption$
%$ Stale under fluctuating conditions$%$ 7heap roust simple and /ell understood for scale-up$
*er+entors
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T$,ical *er+entor
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★ Seed
)ulture,$ 9he seed culture is de!eloped first in the la y the addition of Penicillium chrysogenum spores into a li:uid medium$ When it has
gro/n to the acceptale amount it is inoculated into the
fermenter$
+*$ 9hemedium
is constantly aerated andagitated$
7aronandnitrogen are added sparingly alongside precursor molecules for
penicillin fed-atch style$ 9ypical parameters such as pH temperature
stirrer speed and dissol!ed o(ygen concentration are oser!ed$
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★ Seed
)ulture++$'fter aout 8* hours penicillin egins to e secreted y the fungus$
+$'fter aout B days gro/th is completed the pH rises to .$* or ao!eand penicillin production ceases$
9he Penicillium fungus
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★ .e+oval of
/io+ass+3$Filtration is carried out as ioseparation is re:uired to remo!e the iomass from the culture 2remo!ing the fungus and other impurities
a/ay from the medium /hich contains the penicillin4$
+8$' R otary !acuum filter is commonly employed for filtration as it is
ale to run in continuous mode in any large scale operations$
+)$'fter filtration phosphoric acid
a non-o(idising agent is introduced
to decrease pH from .$* to ;$) so asto pre!ent loss of acti!ity ofpenicillin$
Rotary !acuum filter
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★ Addition of
solvent +;$=rganic sol!ents such as amyl acetateD utyl acetate are added to dissol!e thepenicillin present in the filtrate$
+B$'t this point penicillin is present in thesolution and any other solids /ill e consideredas /aste 2can e used as fertili@ers and animal
feed4$
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★ )entrifugal
traction+.$7entrifugation is done to separate the solid
/aste from the li:uid component /hich contains
the penicillin$
+,$ Esually a dis# centrifuge is used at this point$
*$9he supernatant /ill then e transferred
further in the do/nstream process to continue
/ith e(traction$is# centrifuge - =ne of the most
common type of centrifuge for largescale production
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+$' series of e(traction processes are carried upon the dissol!ed
penicillin to otain a etter purity of the penicillin product$
$9he acetate solution is first mi(ed /ith a phosphate uffer follo/ed
y a chloroform solution and mi(ed again /ith a phosphate uffer and
finally in an ether solution$
3$Penicillin is present in high concentration in the ether solution and
it /ill e mi(ed /ith a solution of sodium icaronate to otain the
penicillin-sodium salt /hich allo/ penicillin to e stored in a stale
po/der form at rtp$
★
traction
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traction8$ 9he penicillin-sodium salt is otained from the li:uid material y as#et centrifugation in /hich solids are easily remo!ed$
1atch e(traction unit 1as#et 7entrifuge" 6(tremely useful in the
remo!al of solids in this case Penicillinsalt
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★ *luid /ed
dr$ing )$ rying is necessary to remo!e anyremaining moisture present in the po/dered
penicillin salt$
;$ In fluid ed drying hot gas is pumped from
the ase of the chamer containing the
po/dered salt inside a !acuum chamer$
B$ Moisture is remo!ed this /ay and this
result in a much drier form of penicillin$
Po/dered penicillin eing
lo/n y hot air
Fluid ed dryingtue
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★
Storage.$ Penicillin is stored in containers and #ept in a drieden!ironment$
9he White Penicillin-Sodium salt
,$9he resulting penicillin 2called Penicillin G4 can e chemically and
en@ymatically modified to ma#e a !ariety of penicillins /ith slightlydifferent properties$
3*$9hese can e semi-synthetic penicillins such as Penicillin
Penicillin = ampicillin and amo(ycillin$
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Increasing ield
+$ 'dding corn syrup$ Selection of strain
3$ Mutation and Selection
8$ Se(ual reproduction
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+$ 7orn syrup 'llo/ ma(imal gro/th of the culture at the e(pense of product
2antiiotic4 formation$ 9his is ecause gro/th and antiiotic
production are in!ersely proportional$More secondary metaolites during stress phase$
Increasing ield
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$ Selection of strain
● Selection of the est strain
depends on the production
rateof the secondary
metaolite 2penicillin4$
% Penicillium notatum 2+ mgDdm34
and Penicillium chrysogenum
2)* mgDdm34$
% Strains are gro/n on cultures in
laoratories and those /ith est
yield is determined$
Increasing ield
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3$ Mutation and Selection9his is ased on trial and chance$
● Se!eral strains of Penicillium are cultured$
● 6thyl methanesulphonate 26MS4 near-ultra!iolet light
in the presence of .-metho(ypsoralen 2.M=P4 are used
as mutagens$
● 7omination of the mutagens leads to a more positi!eresult$
● Strains are e(posed to the mutagens at different intensities andproportions$
● 6ach time the est strains are selected and are further e(posed$
● Statistical tests are done to determine the strains /ith
highest yields$
● ry conidia is stored in silica gel at 8°C at each
Increasing ield
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8$ Se(ual Reproduction
● It /as assumed that the penicillin-producing
mould fungus
Penicillium chrysogenum only reproduced ase(ually through
spores$ Fungus also has a se(ual cycle 2an$. *+34
● 9he progenies possess a comination of genes from oth mating
partners and thus ha!e ne/ properties oth at the molecular le!el
as /ell as in their phenotypes$
% Specific en!ironmental conditions in the dar# under o(ygen
depri!ation conditions in a nutrient medium supplemented /ith
the !itamin iotin$
Increasing ield
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http"DDpenicillin$/i#ispaces$comDGeneralJioprocessJflo/ 2+)D*,D*+34
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http"DDmicroiologyprocesses$logspot$comD*++D+Dpenicillin-production$html2*D*,D*+34
http"DDin!entors$aout$comDodDpstartin!entionsDaDPenicillin$htm2*D*,D*+34
http"DD///$medicinenet$comDpenicillins-oralDarticle$htm 23*D*,D*+34
http"DD///$eolss$netDSample-7haptersD7+BD6;-).-*)-*8$pdf 23D+*D*+34
http"DDmic$sgmCournals$orgDcontentD++3DD*,$full$pdfJhtml 2++D+*D+34
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h ttp"DDarchi!e$io$ed$ac$u#DCdeaconDmicroesDpenicill$htm 2++D+*D+34
http"DD///$sciencedirect$comDscienceDarticleDpiiDS*+8+*,*B**3);*
2*BD++D+34
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