bioteknologi farmasi 1

8/17/2019 BIOTEKNOLOGI FARMASI 1

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BIOTEKNOLOGI FARMASI

‘ISOLASI DAN PURIFIKASI PROTEIN’

DEWI DIANASARI,M.Farm.,Apt.


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PROTEIN

• ‘Protein’ a word derived from Greekword, proteios, which means ‘ of therst rank’ was coined by Jons J.Berzelis ini !"#$ to stress theimportance of this %rops of or%aniccomponds. Protein have been

describeed as the servants of life.&ost proteins are enzymes bt inaddition there also re%latory and

strctral protein.


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• 'el men%andn% riban protein

• Prosedr pemrnian harsmembedakan protein yan%diin%inkan dari protein yan% lain

• 'ifat protein merpakan knci dalammerancan% protokol pemrnian


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Prosedr pemrnian protein dapat dilakkan karena adanya variasi(

)elartan, kran, matan, bindin% specicity *a+nity,sifat hidrofobik-hidrolik


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Nama Jenis Inhibitor Target Protease

PMSF

(Phenylmethylsulfonyl fluoride)

Ireversibel Ser (+ys) !rotease" teri#at!ada !usat a#tif Ser$ys

%en&amidine 'eversibel Ser dan ys !rotease" inhibitor#om!etitif

T* 'eversibel Metalo en&im" helate metals

Pestatin (!enta!e!tida + gugusorgani#)

'eversibel *s!,aidi, !rotease" inhibitor#om!etitif (target" !e!sin- .I/!rotease)

*!rotinin (012mer !e!tida) 'eversibel Serin !rotease- esterase"inhibitor #om!etitif (target"tri!sin- #imotri!sin- !lasmin)

3eu!e!tin (*2leu2leu2*rg) 'eversibel Ser dan ys !rotease" inhibitor#om!etitif

nhibitor Protease( inaktivasi protease yan%dibebaskan selama lisis sel


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Chromatoraph!"M#tho$%

Protein di/er in size and char%e.0hromato%raphic methods takeadvanta%e of both properties.

2 &obile phase2 'tationary phase


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on1e2chan%e 0hromato%raphy *34

• 0ation e2chan%ers contain ne%atively char%ed polymer

• 5nion e2chan%ers contain positively char%ed polymer.

• s e/ected by p6


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Gel 7iltration-'ize 32clsion 0hromato%raphy


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)romato%ra nteraksi

6idrofobik• Protein mempnyai

‘daerah’ hidrofobikdipermkaan yan% dapatberinteraksi den%an resin

• 8esin yan% men%andn%%%s fenil ata alifatikmen%ikat protein( proteinyan% palin% hidrofobikakan terikat palin% kat

• 3lsi dilakkan den%an

pelart yan% kran% polarata den%an menrnkan9%aram:


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3lektroforesis ';'1P5G3

• Pemisahan protein*molekl bermatanberdasarkan kran

dalam medan listrik• Protein didenatrasi

den%an deter%enanion


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5nalisa . )romato%ra ltrasi %el?. )romato%ra anitas

!@A% protein-lar


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T#r!ma &a%!h

bioteknologi farmasi 1

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