The kind of bacteria was identified using Gram’s coloring method. Bacteria was isolated in ½ Luria Berthani (½ LB) medium. The research aim is to isolate β-galactosidase enzyme of thermophile bacteria isolated from Gedong Songo hot water spring, to characterize optimum pH, incubation time, and temperature of the enzyme and to know the specific activity of β-galactosidase in above condition. Suprapti through several qualitative tests concluded that thermophile bacteria isolated from Gedong Songo hot water spring has a potential of β-galactosidase. Β-galactosidase is an enzyme converting lactose to glucose and galactose. Aktivitas spesifik tertinggi pada kondisi optimum tersebut didapatkan pada F4 (60-80 %) yaitu 44,145 Unit/mg protein. Hasil karakterisasi menunjukkan bahwa kondisi optimum enzim β-galaktosidase isolat bakteri termofilik sumber air panas Gedong Songo dengan substrat ONPG adalah pada suhu 63 oC, pH 6,2 dan waktu inkubasi 30 menit. Isolat bakteri termofilik sumber air panas Gedong Songo memiliki sifat gram negatif dan berbentuk batang. Karakterisasi enzim meliputi pH, suhu dan waktu inkubasi optimum dari enzim β-galaktosidase yang diperoleh. Uji aktivitas enzim β-galaktosidase dilakukan dengan menggunakan ONPG sebagai substratnya dan kadar protein diukur dengan menggunakan metode Lowry. Enzim β-galaktosidase diisolasi dengan metode ekstraksi, fraksinasi amonium sulfat bertingkat dan dialisis. Enzim β-galaktosidase diproduksi pada media ½ LB selama 6 jam dengan menggunakan laktosa sebagai penginduksi. Jenis bakteri diidentifikasi dengan metode pewarnaan Gram. Isolasi bakteri menggunakan media ½ Luria Berthani (½ LB). Penelitian ini bertujuan untuk memperoleh enzim β-galaktosidase dari isolat bakteri termofilik sumber air panas Gedong Songo, memperoleh karakter enzim dan menentukan aktivitas spesifik enzim β-galaktosidase pada kondisi optimumnya. Berdasarkan uji kualitatif yang telah dilakukan oleh Suprapti menyatakan bahwa isolat bakteri termofilik sumber air panas Gedong Songo berpotensi menghasilkan enzim β-galaktosidase. RINGKASAN Enzim β-galaktosidase merupakan enzim yang dapat mengubah laktosa menjadi glukosa dan galaktosa.
ENZIM PROTEOLITIK PDF CODE
A similarity analysis based on the 16S rRNA gene sequence showed that IROD3 (Indonesian Red Oncom Day-3) has 99% similarity level with the 16S rRNA gene fragment of Bacillus megaterium strain CS17 (access code Genbank: MG430224.1).ISOLASI DAN KARAKTERISASI ENZIM β-GALAKTOSIDASE DARI ISOLAT BAKTERI TERMOFILIK SUMBER AIR PANAS GEDONG SONGO The result of bacteria isolation was found one isolate which has proteolytic activity in Skim Milk Agar medium which has clear zone diameter of 78.00 mm. The process of Molecular identification process was carried out through analysis of 16S rRNA gene fragment sequences which were amplified using Polymerase Chain Reaction (PCR) method, and continued by sequencing.
Isolation and purification process of bacterial colony was carried out on Nutrient Agar medium with spread technique, production test of protease enzyme was performed using Selective Skim Milk Agar. The purpose of this study was to obtain an isolate of protease-producing bacteria found on post- fermentation oncom 72 hours, and to identify the protease-producing bacteria based on the analysis of 16S rRNA gene. One of effort to increase the production of protease enzymes is looking for new sources of protease particularly from bacterial groups. Protease enzyme has function to hydrolyze peptide bonds in proteins into simpler molecules to digest by the body which important to food industry.
0 kommentar(er)
