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Activity of enzyme fraction was tested using ONPG while protein concentration was measured using Lowry’s method. The β-galactosidase was isolated using extraction, fractionation and dialysis methods. Production of β-galactosidase was done in ½ LB medium for 6 hours using lactose as inducer. 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. IgY and IgG activities were reduced significantly at pH 10.ISOLASI DAN KARAKTERISASI ENZIM β-GALAKTOSIDASE DARI ISOLAT BAKTERI TERMOFILIK SUMBER AIR PANAS GEDONG SONGO Treatment with pepsin significantly reduced IgY and IgG whereas trypsin slightly reduced IgY activities and has no effect on IgG activities.

IgY and IgG activities were significantly reduced at 80✬ and completely destroyed at 90✬. The activities of treated IgY and IgG were tested by enzyme linked immunosorbent assay (ELISA). Commercial antitetanus IgG was used as control. 120).The purified IgY was then treated at different levels of temperatures and pH as well as proteolytic enzymes. IgY was purified by ammonium sulfat precipitation and gel filtration chromatography (Sephadex G. Serum samples were collected 4 weeks following the last immunization. Antitetanus IgY was produced by immunization of chickens with tetanus toxoid, three times weekly at gradual doses of 100, 200, and 300 Lf, respectively.

A study was carried out to find out an alternative method of producing antitetanus antibody (IgY) in chicken and to evaluate its activity at different levels of pH, temperatures and proteolytic enzymes.