Pemurnian dan karakterisasi xilanase ekstraseluler dari streptomyces sp. 234P-16 asal Padang
FAHRURROZI. Purification and Characterization of Extracellular Xylanase from Streptomyces sp. 234P 16 Isolated from Padang.
Abstract
Xylan is the major component of plant hemicellulose and the second most abundant polysaccharide in nature. As a renewable substance, xilan can be degraded as useful end product. Xylanases is a group of enzyme that has ability to hydrolyse hemicellulose into xylan or polymer of xyloses and @ xylooligosaccharides. Complete hydrolysis of xylans involves the synergistic action ofaxylanolytic enzymes, including 13.-xylanase, f!,-xylosidase, :::I: a-glucuronidase, a-L-arabinofuranosidase, and acetyl xylan esterase. ~ Streptomyces sp. 234P-16 produced extracellular xylanase that had unique o characteristics: optimum temperature and pH was 90°C and 5.0 respectively. '2. The purpose of this research was to purify and characterize' the xylanase of III 3 Streptomyces sp. 234P-16. Streptomyces sp. 234P-16 was able to produce two _. xylanolytic enzymes a-L-arabinofuranosidase and (3-xilosidase. Crude extract of '" xylanase was precipitated by 60-70% acetone gradually, was purified with anion ~ exchange chromatography using Sephadex A-50 and gel filtration chromatography using Sephadex G-100. The molecular mass of the purified ~ xylanase after ion-exchange was 42.456 kDa and 16.903 kDa but gel filtration ~ showed the molecular mass 42.456 kDa. The optimum temperature and pH was S. 90°C and 5.0 respectively. The xylanase could resist of heating at 90°C for ~ 180 minute. The result of kinetic analysiS showed that Vmax was 523 :l IJmol/minute/mg and Km was 1.98 mg/ml. Xilan adalah komponen terbesar hemiselulosa tanaman dan merupakan polisakarida terbesar kedua setelah selulosa. Xilan mempunyai potensi yang sangat besar untuk diuraikan menjadi produk akhir yang berguna. Xilanase ~upakan kelompok enzim yang menghidrolisis hemiselulosa dalam hal ini ialah ~n atau polimer dari xilosa dan xilooligosakarida. Pemecahan sempuma xilan nfmerlukan aktivitas sinergis beberapa enzim hidrolitik (hemiselulase), yaitu ePrrlo-1 ,4-r1-xilanase, ~-xilosidase, a-glukuronidase, a-L-arabinofuranosidase, ~n asetil xilan esterase. Streptomyces sp. 234P-16 menghasilkan xilanase etl!;traseluler dengan sifat yang unik yaitu mempunyai suhu optimum 90°C dan pel optimum 5,0. Penelitian ini bertujuan untuk melakukan pemurnian serta kiakterisasi biokimiawi xilanase ekstraseluler yang dihasilkan oleh StrPptomyces sp. 234P-16. Streptomyces sp. 234P-16 dapat memproduksi dua e~im xilanolitik yaitu a-L-arabinofuranosidase dan (3-xilosidase. Xilanase ekstrak kiiar diendapkan dengan 60-70% aseton secara bertahap, dimurnikan dengan k~m atografi penukar anion Sephadex A-50 dan terakhir dimurnikan ~nggunakan kromatografi filtrasi gel Sephadex G-100. Berat molekul xilanase setelah dimumikan dengan penukar anion, yaitu 42,456 dan 16,903 kDa, namun sE!elah dimurnikan dengan filtrasi gel 42,456 kDa. Suhu dan pH optimum 90°C dci1 5,0. Stabil pada suhu 90°C selama 180 menit. Pengukuran terhadap p:!ameter kinetika menghasilkan Vmaks = 523 ~mol/menitlmg dan KIOJ= 1,98 mg/ml
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