Productionofpenicillin

Learningobjective:
Tobeabletoidentifyusefulproductsfrom
microorganisms
Tobeabletoidentifythemicroorganismsusedand
themainstagesintheproductionofpenicillin.
TobeabletodescribehowDownstreamprocessing
iscarriedouttoextractandpurifytheendproduct
offermentation.

UNITII
PENCILLINPRODUCTION

R.KAVITHA,M.PHARM
LECTURER,
DEPARTMENTOFPHARMACUTICS
SRMCOLEGEOFPHARMACY
SRMUNIVERITY

DownstreamProcessing
Productsinafermenterareimpureanddilute,so
needtobepurifiedbydownstreamprocessing.
Thisusuallyinvolvesfiltrationtoseparatethe
microbialcellsfromtheliquidmedium,followedby
chemicalpurificationandconcentrationofthe
product
Downstreamprocessingcanaccountfor50%ofthe
costofaprocess.

 Antibioticsareantimicrobialagentsproduced
naturallybyothermicrobes(usuallyfungior
bacteria).
Thefirstantibioticwasdiscoveredin1896by
ErnestDuchesneand"rediscovered"
byAlexanderFlemmingin1928fromthe
filamentousfungusPeniciliumnotatum.

 Theantibioticsubstance,namedpenicillin,
wasnotpurifieduntilthe1940s(byFlorey
andChain),justintimetobeusedattheend
ofthesecondworldwar.
Penicillinwasthefirstimportantcommercial
productproducedbyanaerobic,submerged
fermentation

 Whenpenicillinwasfirstmadeattheendof
thesecondworldwarusingthefungus
Peniciliumnotatum,theprocessmade1mg
dm3.
Today,usingadifferentspecies(P.
chrysogenum)andabetterextraction
procedurestheyieldis50gdm3.
Thereisaconstantsearchtoimprovethe
yield.

 Antibioticscanbeselectivelytoxicby
targetingsuchfeaturesasthebacterialcell
wall,70Sribosomes,andenzymesthatare
specifictobacteria.
Inthiswaythehumaneukaryoticcellsare
unaffected.

Forexample:

penicillin,ampicillin,amoxycillin,methicillin
Inhibitsenzymesinvolvedinsynthesisof
peptidoglycanforbacterialcellwall,causing
celllysis.
Bacteriocidal
Narrowspectrum littleeffectonGram
negativecells.

OtherantibioticsMOmayaffect:
Cellmembrane
DNAreplication
Transcription
Translation

Antibioticproduction
Thereareover10000differentantibioticsknown,
butonlyabout200incommercial
use,sincemostnewantibioticsarenobetterthan
existingones.
Thereisaconstantsearchfornewantibiotics.
Antibioticsarethemostprescribeddrugsandare
bigbusiness.
Findinganewantibioticandgettingitontothe
marketisaverylongprocessandcantake15years.

AntibioticProduction
Methods

Antibioticsareproducedonanindustrialscaleusinga
varietyoffungiandbacteria.
PenicillinisproducedbythefungusPenicillium
chrysogenumwhichrequireslactose,othersugars,and
asourceofnitrogen(inthiscaseayeastextract)inthe
mediumtogrowwell.
Likeallantibiotics,penicillinisasecondarymetabolite,
soisonlyproducedinthestationaryphase.
Whatsortoffermenterdoesitrequire?
Itrequiresabatchfermenter,andafedbatchprocess
isnormallyusedtoprolongthestationaryperiodand
soincreaseproduction.

 Downstreamprocessingisrelativelyeasy
sincepenicillinissecretedintothe
medium(tokillothercells),sothereisno
needtobreakopenthefungalcells.
However,theproductneedstobevery
pure,sinceitbeingusedasatherapeutic
medicaldrug,soitisdissolvedandthen
precipitatedasapotassiumsaltto
separateitfromothersubstancesinthe
medium.

 Theresultingpenicillin(calledpenicillinG)
canbechemicallyandenzymatically
modifiedtomakeavarietyofpenicillins
withslightlydifferentproperties.
Thesesemisyntheticpenicillins include
penicillinV,penicillinO,ampicillin and
amoxycillin.

1. WhatistheCarbonsource?
lactose
2. Whatisthenitrogensource?
yeast
3. Whatistheenergysource?
glucose
4. Isthefermentationaerobicoranaerobic?
aerobic
5. Whatistheoptimumtemperature?
25 - 27C
6. Ispenicillinaprimaryorsecondarymetabolite?
secondary
7. Whatvolumefermenterisused?
3
8. Whyisn'talargerfermenterused? 40 200 dm
To difficult to aerate
9. Whenispenicillinproduced?
10. Howlongcanitbeproducedfor? 40 hours after main increase in fungal mass
11. Whatwasthefirstfungusknowntoproducepenicillin?
140 hours (180 40 hours)
12. Whatspeciesproducesabout60mg/dm3ofpenicillin?
Penicillin notatum
13. Howdidscientistsimprovetheyieldstillfurther?
Penicillin chrysogenum
14. Whatisthesubstrate?
Genetic modification
15. Whyisbatchcultureused?
Corn steep liquor
16. Whataretheprocessesinvolvedindownstreamprocessing?
Secondary metabolite
a)
b)
c) Filtration of liquid
Extraction from filtrate by counter current of butylacetate
17. Whycan'tpenicillinbetakenorally?
18. Nametheformofpenicillinwhichcanbetakenorally.
Precipitation by potassium salts
19. HowdoesPenicillinkillbacteria?
Destroyed by stomach acid
20. WhyareGramnegativebacterianotkilledbypenicillin?
Penicillin V, ampicillin
Stops production of cell wall
Different cell wall