CPT Presentation

8/17/2019 CPT Presentation

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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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.eferenceshttp"DDpenicillin$/i#ispaces$comDHome 2+)D*,D*+34

http"DDpenicillin$/i#ispaces$comDGeneralJioprocessJflo/ 2+)D*,D*+34

http"DDpenicillin$/i#ispaces$comDariousJtypesJofJPenicillin 2+)D*,D*+34

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

http"DD///$sciencedaily$comDreleasesD*+3D*+D+3*+*.*.8+3.$htm 2++D+*D+34

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

http://penicillin.wikispaces.com/Homehttp://penicillin.wikispaces.com/Homehttp://penicillin.wikispaces.com/General+bioprocess+flowhttp://penicillin.wikispaces.com/General+bioprocess+flowhttp://penicillin.wikispaces.com/Various+types+of+Penicillinhttp://penicillin.wikispaces.com/Various+types+of+Penicillinhttp://microbiologyprocesses.blogspot.com/2011/12/penicillin-production.htmlhttp://inventors.about.com/od/pstartinventions/a/Penicillin.htmhttp://www.medicinenet.com/penicillins-oral/article.htmhttp://www.eolss.net/Sample-Chapters/C17/E6-58-05-04.pdfhttp://www.eolss.net/Sample-Chapters/C17/E6-58-05-04.pdfhttp://mic.sgmjournals.org/content/113/2/209.full.pdf+htmlhttp://mic.sgmjournals.org/content/113/2/209.full.pdf+htmlhttp://www.sciencedaily.com/releases/2013/01/130108084138.htmhttp://archive.bio.ed.ac.uk/jdeacon/microbes/penicill.htmhttp://archive.bio.ed.ac.uk/jdeacon/microbes/penicill.htmhttp://www.sciencedirect.com/science/article/pii/S0141022907003560http://www.sciencedirect.com/science/article/pii/S0141022907003560http://archive.bio.ed.ac.uk/jdeacon/microbes/penicill.htmhttp://archive.bio.ed.ac.uk/jdeacon/microbes/penicill.htmhttp://www.sciencedaily.com/releases/2013/01/130108084138.htmhttp://mic.sgmjournals.org/content/113/2/209.full.pdf+htmlhttp://mic.sgmjournals.org/content/113/2/209.full.pdf+htmlhttp://www.eolss.net/Sample-Chapters/C17/E6-58-05-04.pdfhttp://www.medicinenet.com/penicillins-oral/article.htmhttp://inventors.about.com/od/pstartinventions/a/Penicillin.htmhttp://microbiologyprocesses.blogspot.com/2011/12/penicillin-production.htmlhttp://penicillin.wikispaces.com/Various+types+of+Penicillinhttp://penicillin.wikispaces.com/Various+types+of+Penicillinhttp://penicillin.wikispaces.com/General+bioprocess+flowhttp://penicillin.wikispaces.com/General+bioprocess+flowhttp://penicillin.wikispaces.com/Home


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