Purification and characterization of Brevibacterium sp. amylase
Purifikasi dan karakterisasi enzim amilase dari Brevibacterium sp
dc.contributor.advisor | Meryandini, Anja | |
dc.contributor.advisor | Yopi | |
dc.contributor.author | Putranto, Widyo Tri | |
dc.date.accessioned | 2013-02-12T03:19:07Z | |
dc.date.available | 2013-02-12T03:19:07Z | |
dc.date.issued | 2012 | |
dc.identifier.uri | http://repository.ipb.ac.id/handle/123456789/60687 | |
dc.description.abstract | Amylase is an enzyme that used widely in various industrial field. This enzyme specifically catalyze the hydrolysis of -1,4-glycosidic bond on starch. To fulfill industrial need, the search for a new promising source of amylase is important. One of amylase source that has not been well explored yet is marine microbe. The purpose of this research was to purify and characterize the amylase from Brevibacterium sp. isolated from Jakarta Bay. The Brevibacterium sp. was cultured in Artificial Sea Water (ASW) medium. Measurement of specific amylase activity was performed by modified Miller method. The highest crude enzyme activity was produced 96 hours after the cultivation which is reaching 1,2 U/mL. After precipitation using ammonium sulphate, the crude enzyme solution were purified by gel filtration chromatography. SDS-polyacrylamide analysis result showed that the molecular weight of the enzymes were 79 kDa, 55,7 kDa, and 33,6 kDa. The partial purified enzyme has optimum activity on temperature 50oC and pH 6,4. | en |
dc.description.abstract | Amilase adalah enzim yang digunakan di berbagai bidang industri. Enzim ini mengkatalisis hidrolisis dari ikatan 1,4-glikosidik pada pati. Untuk memenuhi kebutuhan industri, pencarian terhadap sumber amilase baru yang menjanjikan sangat penting. Salah satu sumber amilase yang belum banyak dieksplorasi adalah mikroba laut. Penelitian ini bertujuan mempurifikasi dan mengkarakterisasi enzim amilase dari Brevibacterium sp., yang diisolasi dari perairan Teluk Jakarta. Isolat Brevibacterium sp. dikulturkan di media Artificial Sea Water (ASW). Pengukuran aktivitas amilase dilakukan menggunakan metode Miller yang dimodifikasi. Aktivitas amilase tertinggi pada enzim ekstrak kasar diproduksi pada jam ke-96 setelah pengkulturan, yaitu mencapai 1,2 U/mL. Setelah presipitasi dengan menggunakan ammonium sulfat, larutan enzim ekstrak kasar dimurnikan dengan kromatografi gel filtrasi. Hasil analisis SDS-PAGE menunjukkan berat molekul protein adalah 79 kDa, 55,7 kDa, dan 33,6 kDa. Berdasarkan karakterisasi, enzim bekerja optimum pada suhu 50oC dan pH 6,4. | |
dc.publisher | IPB ( Bogor Agricultural University ) | |
dc.subject | amylase | en |
dc.subject | purification | en |
dc.subject | characterization | en |
dc.subject | Brevibacterium | en |
dc.title | Purification and characterization of Brevibacterium sp. amylase | en |
dc.title | Purifikasi dan karakterisasi enzim amilase dari Brevibacterium sp |
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