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KARAKTERISASI ENZIM AMILASE DARI BAKTERI Bacillus amyloliquefaciens Ningsih, Dian Riana; Rastuti, Undri; Kamaludin, Ridlwan
Prosiding Vol 3, No 1 (2012)
Publisher : Prosiding

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Abstract

Enzim Amilase merupakan salah satu jenis enzim yang berperan penting dalam industri. Enzim amilase digunakan untuk menghidrolisis pati menjadi molekul karbohidrat yang lebih sederhana, yaitu dekstrin, maltosa dan glukosa. Industri yang menggunakan amilase antara lain: dalam industri kertas, industry detergen, industry tekstil, industry obat dan industri roti dan kue. Bacillus amyloliquefacien merupakan salah satu bakteri yang dapat menghasilkan amilase. Tujuan dari penelitian ini adalah menentukan aktivitas enzim amylase dan mengkarakterisasi sifat biokimia enzim amylase dari B. amyloliquefaciens. Tahapan penelitian ini adalah penentuan waktu produksi optimum enzim amylase, produksi amylase dan penentuan aktivitas enzim amylase pada berbagai suhu dan pH. Penentuan aktivitas amylase menggunakan metode Nelson Somogyi. Hasil penelitian menunjukkan bahwa enzim amylase yang dihasilkan oleh B. amyloliquefaciens mempunyai waktu produksi optimum pada jam ke 24 (1.4986 U/ml), temperature optimum 30-60 oC dan pH optimum 6-7.
Hidrolisis Pati Ganyong (Canna edulis) dengan Amilase Bakteri Flavobacterium sp. PTBT I untuk Produksi Bioetanol Ningsih, Dian Riana; Zusfahair, Zusfahair; Fatoni, Amin
Jurnal Natur Indonesia Vol 15, No 2 (2013)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (205.385 KB) | DOI: 10.31258/jnat.15.2.92-98

Abstract

Bioethanol is an alternative energy of fuels produced from vegetable materials. Vegetable materials that can be used as rawmaterial for bioethanol is ganyong because it contains 22.60 g starch in 100 g ganyong. The production of bioethanol fromstarch material consisted of two steps, hydrolysis and fermentation. One of the steps to increase the value of bioethanolfrom starch of ganyong was hydrolysis process using thermostable amylase enzyme isolated from Flavoacterium sp.PTBT I bacteria was isolated from hot spring of Pancuran Tujuh Baturraden. The aim of this research was to use thermostableamylase to hydrolyze starch of ganyong and glucose produced to result bioethanol. The result of this research showed thatthe optimum condition hydrolysis starch of ganyong was using thermostable amylase acquired at substrate concentrationof 3% (b/v), and incubation time of about 75 minutes. The value of bioethanol increased with time of fermentation, from thefirst to fourth day, which was 0.8361; 2.2379; 5.7590 and 10.5787% (v/v), respectively.
Pemurnian Parsial dan Karakterisasi Urease dari Biji Kacang Panjang (Vigna unguiculata subsp sesquipedalis L.) Zusfahair, Zusfahair; Ningsih, Dian Riana; Fatoni, Amin; Pertiwi, Darul Santri
ALCHEMY Jurnal Penelitian Kimia Vol 14, No 1 (2018): Alchemy Jurnal Penelitian Kimia
Publisher : UNIVERSITAS SEBELAS MARET (UNS)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20961/alchemy.14.1.13000.72-83

Abstract

Urease merupakam enzim yang digunakan dalam hidrolisis urea menjadi amoniak dan asam bikarbonat dan telah banyak digunakan dalam proses industri. Tujuan penelitian adalah isolasi dan pemurnian urease dari kacang panjang serta karakterisasinya. Penelitian dimulai dengan melakukan perkecambahan biji kacang panjang selama 8 hari. Kecambah biji kacang panjang selanjutnya diekstraksi dengan menggunakan buffer fosfat pH 7 dan dipisahkan menggunakan sentrifugasi sehingga diperoleh ekstrak kasar urease. Ekstrak kasar urease selanjutnya difraksinasi menggunakan aseton pada tingkat konsentrasi 20, 40, 60 dan 80%. Fraksi yang mempunyai aktivitas spesifik paling tinggi selanjutnya dianalisis menggunakan metode SDS-PAGE untuk menentukan berat molekulnya dan dikarakterisasi lanjut meliputi: pengaruh suhu, pH, konsentrasi substrat dan penambahan ion logam terhadap aktivitas urease. Aktivitas urease ditentukan dengan metode Nessler. Hasil penelitian menunjukkan aktivitas spesifik urease dari kacang panjang paling tinggi ditemukan pada fraksi aseton (FA) 20. Hasil analisis berat molekul dengan metode SDS-PAGE diperoleh beberapa pita protein yang diduga berukuran sekitar 25 KDa dan 17 KDa. Kondisi optimum dari aktivitas urease diperoleh pada suhu 30 ºC, pH 7 dan konsentrasi urea 16,6 mM dengan nilai aktivitas 407,62 U/mL. EDTA dan ion logam dalam CaCl2, NaCl, NiCl2 dan CuCl2 pada variasi konsentrasi 10-3, 10-4  dan 10-5 M merupakan inhibitor urease FA 20 dari kacang panjang.Partial Purification and Characterization of Urease from Asparagus Bean (Vigna unguiculata subsp sesquipedalis L.). Urease is an enzyme used in urea hydrolysis to ammonia and bicarbonate acid and has been widely used in industrial processes. The study focused on isolation and purification of urease from asparagus beans and its characterization. The study was started with germination of asparagus beans for 8 days. Germinated asparagus beans were further extracted using phosphate buffer pH 7 and separated by centrifugation to obtain a crude extract of urease. The crude extract of urease was further fractionated using acetone at concentrations of 20, 40, 60 and 80%. The fraction with highest specific activity was then analyzed using SDS-PAGE method to determine its molecule weight and characterized further including the influence of temperature, pH, substrate concentration, and metal ion addition to urease activity. The urease activity was determined by the Nessler̕ s method. The results showed that the specific activity of urease from asparagus beans was found with highest activity in fraction of acetone (FA) 20. Analytical result using SDS-PAGE method was obtained some protein bands having molecular weights about 25 KD and 17 KDa. The optimum conditions of urease activity was obtained at 30 °C, pH 7, incubation time 20 min and urea concentration 16.6 mM with activity value 407.62 U/mL. EDTA and metal ions contained in CaCl2, NaCl, NiCl2 and CuCl2 at concentrations of 10-3, 10-4 and 10-5 M were FA 20 urease inhibitors.
Determination of Cu and Pb concentrations based on urease activity inhibition of Durio zibethinus L. seeds Zusfahair, Zusfahair; Fatoni, Amin; Ningsih, Dian Riana; Riapanitra, Anung
Molekul Vol 16, No 2 (2021)
Publisher : Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (217.71 KB) | DOI: 10.20884/1.jm.2021.16.2.736

Abstract

The determination of heavy metal concentrations has been carried out using sophisticated instruments, and therefore a simple and reliable alternative method is needed as a comparison. The study aimed to determine Cu and Pb concentration of standard solution using the urease activity inhibition method of Durio zibethinus L.  seeds.  The research started with urease extraction from Durio D. zibethinus L. seeds. The activity of the obtained extract was determined using the Nessler method. The optimum substrate concentration was also determined. Urease activity inhibition was carried out using various metal solution concentrations, which continued by plotting a log graph of urea concentration vs. %inhibition. The obtained graph would then determine the metal concentration in a synthetic water sample. The data was then compared to the measurement, determined by the Atomic Absorption Spectrophotometry (AAS) method. Results of the study showed that the urease activity of D. zibethinus L.seeds was 296.774 U/mL. Urease activity was optimum at a urea concentration of 0.3 M. The comparison Cu, and Pb concentration determination using the urease inhibitory activity and AAS methods showed no significant difference at 95% confidence level. This research showed that urease of D. zibethinus L. seed could be used to determine Cu and Pb's concentration based on its inhibiting activity.