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PEMURNIAN DAN KARAKTERISASI ENZIM ASETIL-COA HIDROLASE

PEMURNIAN DAN KARAKTERISASI ENZIM ASETIL-COA HIDROLASE. Donny Nugraha M 20508024. PENDAHULUAN. MEKANISME REAKSI. AGENDA. TEKNIK ANALISIS. PENGGUNAAN. KESIMPULAN. PENDAHULUAN. Senyawa yang dioksidasi siklus Krebs. ..?. ASETIL- CoA.

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PEMURNIAN DAN KARAKTERISASI ENZIM ASETIL-COA HIDROLASE

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  1. PEMURNIAN DAN KARAKTERISASI ENZIM ASETIL-COA HIDROLASE Donny Nugraha M 20508024

  2. PENDAHULUAN • MEKANISME REAKSI AGENDA • TEKNIK ANALISIS • PENGGUNAAN • KESIMPULAN

  3. PENDAHULUAN Senyawa yang dioksidasisiklus Krebs ..? ASETIL-CoA “As donor substrats in co-translational, post-translational or chemical modification : acetylation”

  4. PENDAHULUAN • NC-IUBMB3.1.2.1 • Acetyl-CoAacylase, acetyl-CoAthiol esterase, acetyl-CoAdeacylase, Ach1 • Pertamaditemukandihatibabi (1952) • Hati & otakTikus, Jaringanadiposa Hamster, Saccharomyces cerevisiae

  5. MEKANISMEREAKSI MengkatalisisreaksihidrolisisAsetil-CoAmenjadiAsetatdanCoA

  6. TEKNIKANALISIS Gel Filtration chromatography DEAE – Sepharose SDS-PAGE (Sodium DodecylSulphate- PolyAcrilamide-Gel Electrophoresis)

  7. TEKNIKANALISIS Homogenity and Molecular Properties Menggunakan prosedur kromatografi DEAE-Sepharose, Sepharose CL-4B, Hidroxilapatit Enzim terdiri kira-kira 0,2 % dari total “Cellular protein” Molecular Weight Analysis Menggunakan elektroforesis jel - metode Laemmli dan dibandingkan dengan berat molekul standar (miosin 205kDa, Bovin serum albumin 66kDa) Mr 64,000 ± 5,000 . monomeric

  8. TEKNIKANALISIS Isoelectric Analysis Pita “ Single protein” padapI=5,8 pH Dependence Analysis Pengujian pH dari 4 Sampai 10

  9. TEKNIKANALISIS Temperature Optimum Analysis Aktivitasenzimdari 5 sampai 65C ,maksimumterjadipada 37 -42C Amino Acid Analysis

  10. TEKNIKANALISIS Substrat Specificity Analysis

  11. PENGGUNAAN Pharmaceutical Uses Enzyme would inhibit acetylation of platelets, thus increasing clotting time Agriculture Uses

  12. KESIMPULAN • Mr 64,000 ± 5000 Da • Isoelectric point 5,8 • pH optimum 8,0 • Temperatur Optimum 37 – 42C • Irreversibeldenaturasisetelah 1 menitpada 70C • Substrattergantungspesifisitas

  13. SEKIAN & TERIMAKASIH

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