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All Journal CAKRA KIMIA (Indonesian E-Journal of Applied Chemistry) INDONESIAN JOURNAL OF LEGAL AND FORENSIC SCIENCES Journal Pharmaceutical Science and Application (JPSA) Jurnal Kimia (Journal of Chemistry) METAMORFOSA Journal of Biological Sciences Buletin Udayana Mengabdi Jurnal Farmasi Udayana Jurnal Veteriner ANNALES BOGORIENSES JFIOnline
I Nengah Wirajana
Program Studi Kimia, Fakultas Sains Dan Matematika, Universitas Kristen Satya Wacana
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Dentistry Education Electrical & Electronics Engineering Environmental Science Health Professions Immunology & microbiology Law, Crime, Criminology & Criminal Justice Materials Science & Nanotechnology Medicine & Pharmacology Public Health Veterinary

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HIDROLISIS BATANG JAGUNG SECARA ENZIMATIK DARI TANAH HUTAN MANGROVE Ni Nengah Kartini Asih; Putu Suarya; Ida Bagus Putra Manuaba; I Nengah Wirajana
CAKRA KIMIA (Indonesian E-Journal of Applied Chemistry) Vol 6 No 2 (2018): Volume 6, Nomor 2, 2018
Publisher : Magister Program of Applied Chemistry, Udayana University, Bali-INDONESIA

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ABSTRAK: Tanah hutan mangrove merupakan salah satu sumber selulase. Aktivitas selulase pada tanah hutan mangrove pantai suwung kauh dengan substrat sekam padi telah dilakukan pada penelitian sebelumnya. Tujuan penelitian ini adalah untuk mengetahui waktu hidrolisis optimum dan aktivitas selulase dari tanah hutan mangrove dengan substrat batang jagung tanpa dan dengan delignifikasi. Tanah hutan mangrove Pantai Suwung Kauh Denpasar Bali digunakan secara langsung sebagai sumber selulase. Batang jagung tanpa dan dengan delignifikasi masing – masing dicampur dengan tanah hutan mangrove dan diinkubsi pada suhu 30oC dan pH 7,0 dengan variasi waktu inkubasi 0, 3, 5, 7, 9, dan 11 hari. Gula pereduksi hasil hidrolisis ditentukan dengan menggunakan metode Nelson – Somogyi yang absorbansinya diukur dengan spektrofotometer UV-Vis pada panjang gelombang 540 nm. Aktivitas selulase ditentukan berdasarkan penambahan produk gula pereduksi yang dihasilkan dalam rentang waktu inkubasi. Hasil penelitian menunjukkan bahwa waktu hidrolisis optimum pada sampel tanpa delignifikasi terjadi pada waktu inkubasi selama 5 hari, dengan konsentrasi gula pereduksi 4.5285 mg/mL. Sedangkan, waktu hidrolisis optimum pada sampel dengan delignifikasi terjadi pada waktu inkubasi selama 3 hari (B1), dengan konsentrasi gula pereduksi 16.2340 mg/mL. Aktivitas selulase tertinggi pada sampel tanpa delignifikasi dari hari ke-3 sampai ke-5 sebesar 2.6729 U/mL; dan pada sampel delignifikasi dari hari ke-1 sampai ke-3 sebesar 5.4328 U/mL. Hasil ini mengindikasikan bahwa tanah hutan mangrove memiliki aktivitas selulase untuk menghidrolisis substrat batang jagung dan proses delignifikasi berpengaruh terhadap aktivitas selulase. Kata kunci : batang jagung, delignifikasi, tanah hutan mangrove, selulase ABSTRACT: Mangrove forest soil is one of the sources of cellulase. Cellulase activity in mangrove forest soil from coast of Suwung Kauh Denpasar Bali with rice husk substrate has been carried out in previous studies. The purpose of this study is to determine the optimal hydrolysis time and cellulase activity of mangrove forest soil with corn stalks substrate with and without delignification. The mangrove forest soil from coast of Suwung Kauh Denpasar Bali was directly as a source of cellulase. The corn stalks with and without delignification were mixed with mangrove forest soil and incubated at 30oC and pH 7.0 with incubation times of 0, 3, 5, 7, 9 and 11 days, respectively. The reducing sugar of hydrolysis results was determined by using the Nelson-Somogyi method that the absorbances were measured by the spectrophotometer UV-Vis at wavelength 540 nm. The cellulase activities were determined based on the concentration of reducing sugar that resulted in the incubation period. The results showed that the optimum hydrolysis time in the sample without delignification occurred at an incubation time of 5 days, with a concentration of reducing sugar 4.5285 mg/mL. Whereas, the optimum hydrolysis time in the sample with delignification occurred at the incubation time for 3 days, and with a concentration of reducing sugar 16.2340 mg/mL. The highest cellulase activity in the sample without delignification was from days 3 to 5 of 2.6729 U / mL; and in the delignification sample was from days 1 to 3 of 5,4328 U / mL. The results of this study were that mangrove forest soil had the cellulase activity to hydrolyze substrate and delignification process had a effect on the cellulase activity.
VARIASI KONSENTRASI BUAH ASAM (Tamarindus indica L.) DAN SUSU SKIM TERHADAP KUALITAS YOGHURT KUNIR ASAM Ni Putu Rahayu Artini; Ida Bagus Putra Manuaba; I Nengah Wirajana
CAKRA KIMIA (Indonesian E-Journal of Applied Chemistry) Vol 3 No 3 (2015)
Publisher : Magister Program of Applied Chemistry, Udayana University, Bali-INDONESIA

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Abstract

ABSTRAK: Penelitian ini bertujuan untuk mengetahui pengaruh variasi konsentrasi buah asam (Tamarindus indica L.) dan  susu skim untuk menghasilkan kualitas yoghurt sesuai  dengan SNI 01-2981-2009.Rancangan percobaan yang dilakukan dalam penelitian ini adalah Rancangan Acak Lengkap (RAL) yang terdiri atas sembilan perlakuan. Yoghurt kunir asam dibuat dari variasi penambahan variasi konsentrasi Tamarindus indica L. 30%, 40%, dan 50% (b/V) dan  susu skim 5%, 10%, dan 15% (b/V). Sifat fisika, kimia, dan mikrobiologi  yoghurt kunir asam diamati. Dihasilkan kualitas terbaik yoghurt kunir asam dengan penambahan 30% Tamarindus indica L. (b/V0dan 15% susu skim (b/V). Dengan hasil analisis penampakan cairan kental; konsistensi homogen; rasa asam; bau khas; viskositas 89,3 cP; pH 4,85; kadar abu 1,52%; kadar lemak total 2,53%; kadar protein total 3,74%; kadar asam laktat 0,223%, kadar kurkumin 0,389%; cemaran logam Pb dan Cu serta Total Coliform dan E. coli negatif.ABSTRACT:.The objective of this research was to determinethe influence of concentrated Tamarindus indica L. and skim milk powder in producing tumuric curcumin yogurt towards its product based on SNI 01-2981-2009. The research was conducted in completely randomized design which consisted of nine treatments. The yogurt mixtures were made from a variation of 30%, 40%, and 50% of Tamarindus indica L. and addition of  5%, 10%, and 15% of skim milk powder.  Physical, chemical, and microbiology properties of the turmeric curcuma yogurts were observed.  The results showed the best quality of turmeric curcumin  yogurt was formulated by the addition of 30% Tamarindus indica L. and 15% skim milk powder,  with the results of the analysis: the appearance of a viscous fluid; homogeneous consistency; sour taste; distinctive smell; viscosity of 89.3 cP; pH of 4.85; ash content of 1.52%; total fat content of 2.53%; total protein content of 3.74%; lactic acid levels of 0.22%, curcumin content of 0,389%; however the Pb, Cu, Coliform and E. coli were not detected.
ANALISIS POTENSI PROTEASE EKTRASELULER TANAH HUTAN MANGROVE PANTAI SUWUNG KAUH BALI Inten Hardianti Nizar; I Nengah Wirajana; A.A.I.A Mayun Laksmiwati
CAKRA KIMIA (Indonesian E-Journal of Applied Chemistry) Vol 3 No 3 (2015)
Publisher : Magister Program of Applied Chemistry, Udayana University, Bali-INDONESIA

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Abstract

ABSTRAK: Potensi tanah hutan mangrove pantai Suwung Kauh Bali sebagai sumber protease dapat diketahui dengan melakukan uji aktivitas protease ekstraseluler. Pada penelitian ini telah dilakukan pengukuran aktivitas protease ekstraseluler dan penentuan pengaruh waktu inkubasi serta penambahan toluena terhadap aktivitas protease. Sampel yang digunakan sebagai sumber enzim berupa slurry dan direaksikan dengan substrat kasein 0,3% selama 3,6,9 dan 24 jam dengan dan tanpa penambahan toluena 1% (v/v). Produk reaksi enzimatis diukur dengan metode kolorimetri. Aktivitas protease tertinggi yang diperoleh sebesar 1,9 x 10-4 U/mL dengan penambahan toluena pada waktu inkubasi 6 jam dan sebesar 1,2 x 10-4 U/mL tanpa penambahan toluena pada waktu inkubasi 9 jam. Hasil ini menunjukkan bahwa protease ekstraseluler pada tanah hutan mangrove yang dihasilkan oleh mikroba proteolitik memilki potensi digunakan untuk eksplorasi enzim. Waktu inkubasi dan penambahan toluena tidak berpengaruh signifikan terhadap aktivitas protease.   ABSTRACT: The potency of mangrove soil in Suwung Kauh Bali as a source of protease has been determined by protease activity assay. This research has been done to determine protease activity and the effect of incubation time and the addition of toluene to the protease activity. The slurry of soil was used as a source of extracellular  enzyme for protease assay, which was reacted with casein 0,3% for 3, 6, 9, and 24 hours with and without the addition of toluene 1% (v/v). The enzymatic reaction product was measured by colorimetric method. The highest protease activity with addition of toluene was 1,9 x 10-4 U/mL at 6 hours incubation and without toluene was 1,2 x 10-4 U/mL at 9 hours incubation. These results showed extracelluler protease on mangrove soil produced by proteolytic microorganisms had a potency to be used in enzyme exploration. Furthermore, the incubation time and addition of toluene had no significant effect to protease activity.
PENGARUH TOLUENA DAN WAKTU INKUBASI TERHADAP AKTIVITAS SELULASE DARI TANAH HUTAN MANGROVE Ni Komang Lia Wahyuni; I Nengah Wirajana; I Wayan Budiaarsa Suyasa
CAKRA KIMIA (Indonesian E-Journal of Applied Chemistry) Vol 2 No 2 (2014)
Publisher : Magister Program of Applied Chemistry, Udayana University, Bali-INDONESIA

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Abstract

 ABSTRAK: Tanah hutan mangrove diketahui memiliki biodiversitas yang tinggi sebagai lokasi yang berpotensi untuk eksplorasi enzim. Salah satu enzim yang dapat dieksplorasi dari tanah hutan mangrove adalah selulase yang merupakan biokatalisator yang banyak digunakan dalam bidang industri. Tidak seperti pengukuran aktivitas selulase murni atau ekstrak kasar yang berasal dari salah satu sumbernya, pengukuran aktivitas selulase secara langsung dari tanah sering mengalami kesulitan dan banyak faktor yang harus dipelajari. Penelitian ini bertujuan untuk mengetahui pengaruh penambahan toluena dan waktu inkubasi reaksi enzimatis terhadap aktivitas selulase yang terdapat pada tanah hutan mangrove pantai Suwung Bali. Pengukuran aktivitas selulase dilakukan dengan metode CMC (Carboxymethyl Cellulose Assay) pada sampel tanah (slurry, pelet, dan supernatan) dengan dan tanpa penambahan toluena dengan waktu inkubasi reaksi enzimatis 1 dan 24 jam. Glukosa yang dihasilkan dari reaksi dengan substrat CMC (Carboxymethyl Cellulose) dianalisis secara spektrofotometri UV-Vis pada panjang gelombang 540 nm setelah direaksikan dengan asam 3,5-dinitrosalisilat (DNS). Hasil penelitian ini menunjukkan bahwa penambahan antiseptik toluena dan waktu inkubasi reaksi enzimatis berpengaruh terhadap aktivitas selulase tanah hutan mangrove pantai Suwung Bali. Aktivitas selulase tertinggi sebesar 249,26 U/mL diperoleh pada lumpur dengan penambahan toluena dan inkubasi reaksi enzimatis 1 jam. ABSTRACT: Mangrove soil has high biodiversity and has been well known as potential location for enzymes exploration. One of the enzymes explored from mangrove soil is cellulase which is a biocatalysator commonly used in industries. Unlike the measurement of cellulase activity of pure or crude extract obtained from one source, direct measurement of cellulase activity of the soil often counter many obstacles and many factors are involved that need to be elaborated. The aim of this study is to determine the effects of toluene addition and incubation time of enzymatic reaction on activity of cellulase existing on soil of mangrove forest of Suwung Bali coastal. The measurement of cellulase activity was conducted by the method of CMC (Carboxymethyl Cellulose Assay) on soil samples (slurry, pellet, and supernatant) with and without the addition of toluene to the enzymatic reaction incubation time 1 and 24 hours. Glucose produced from reaction with the substrate CMC (Carboxymethyl Cellulose) was analyzed by UV-Vis spectrophotometer at a wavelength of 540 nm (? 540 nm). The results of this study showed that the addition of toluene antiseptic and incubation time of enzymatic reaction influence the activity of cellulase existing on soil of mangrove forest of Bali Suwung coastal. The highest cellulase activity was 249.26 U/mL obtained on slurry with the addition of toluene and with 1-hour incubation of the enzymatic reaction.
DETEKSI MUTASI KODON 510 dan 511 DAERAH RRDR GEN rpoB PADA ISOLAT KLINIK Mycobacterium tuberculosis MULTIDRUG RESISTANT DI BALI DENGAN PCR-RESTRICTION FRAGMENT LENGHT POLYMORPHISM Made Rai Dwitya Wiradiputra; Sagung Chandra Yowani; I Nengah Wirajana
CAKRA KIMIA (Indonesian E-Journal of Applied Chemistry) Vol 4 No 2 (2016)
Publisher : Magister Program of Applied Chemistry, Udayana University, Bali-INDONESIA

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Abstract

ABSTRAK: Tujuan penelitian ini adalah untuk melakukan deteksi mutasi daerah RRDR gen rpoB Mycobacterium tuberculosis khususnya pada kodon 510 dan 511 dari isolat klinis Multidrug Resistant Tuberculosis (MDR-TB) di Bali dengan metode Polymerase Chain Reaction – Restriction Fragment Length Polymorphism (PCR-RFLP). Isolat M. tuberculosis H37Rv digunakan sebagai kontrol bakteri yang tidak mengalami mutasi dan empat isolat klinis MDR-TB digunakan sebagai sampel pada penelitian ini. Proses PCR-RFLP meliputi dua tahap, yaitu amplifikasi (PCR) dan digesti. Produk PCR hasil amplifikasi didigesti dengan enzim PvuII (New England Biolabs) melalui proses inkubasi pada suhu 37oC selama 3 jam diikuti dengan inaktivasi ice shock pada suhu -20oC selama 5 menit. Hasil penelitian ini menunjukan bahwa enzim restriksi PvuII dapat mendeteksi mutasi kodon 510 dan 511 daerah RRDR gen rpoB M. tuberculosis dengan teknik PCR-RFLP. Pada isolat 134 diketahui terdapat mutasi pada kodon 510 dan/atau 511 sedangkan pada isolat P10, P11, dan P16 tidak ditemukan adanya mutasi pada kodon 510 dan 511. Berdasarkan hasil penelitian sebelumnya, diketahui pula bahwa mutasi yang terjadi pada isolat 134 adalah mutasi kodon 510 (CAG?CTG).   ABSTRACT: The objective of this study is to detect mutation in the region of RRDR of rpoB gene Mycobacterium tuberculosis particularly at codon 510 and 511 from MDR-TB clinical isolates in Bali using Polymerase Chain Reaction – Restriction Fragment Length Polymorphism (PCR-RFLP) method. Isolate of M. tuberculosis H37Rv was used as control of non-mutated bacteria, and four MDR-TB clinical isolates were used for sample in this study. PCR-RFLP was conducted in two steps which were amplification (PCR) and digestion. PCR products were digested using PvuII restriction enzyme (New England Biolabs) through incubation at 37oC for 3 hours followed by ice shock inactivation at -20oC for 5 minutes. The result of this study showed that PvuII restriction enzyme could detect mutation of codon 510 and 511 in the region of RRDR of rpoB gene M. tuberculosis using PCR-RFLP. In isolate 134, mutation at codon 510 and/or 511 was found while there were no mutation of codon 510 and 511 detected in isolate P10, P11, and P16. Based on previous research, it was found that the mutation occurred in isolate 134 was at codon 510 (CAG?CTG).
SUHU DAN WAKTU OPTIMUM PROSES EKSTRAKSI ANTOSIANIN DALAM UBI JALAR UNGU (Ipomoea batatas L.) DENGAN ?-L-ARABINOFURANOSIDASE I N. Wirajana; N. M. T. Juliasari; A.A. I.A.M. Laksmiwati; N. W. Bogoriani
Jurnal Kimia (Journal of Chemistry) Vol.13 No.1 Januari 2019
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (476.09 KB) | DOI: 10.24843/JCHEM.2019.v13.i01.p14

Abstract

Enzyme-assisted extraction (EAE) method is one of the most environmentally friendly methods of enzyme application in the extraction of bioactive compounds. The purpose of this study was to determine the optimum temperature and time required in the extraction of anthocyanin compounds from purple sweet potato (Ipomoea batatas L.) with and without ?-L-arabinofuranosidase (AbfA) - assisted. The AbfA enzyme was obtained from Saccharomyces cerevisiae recombinant strain BJ1824 contain pYHMI-Af plasmid. The optimum temperature and time in the extraction of anthocyanin compound with and without ?-L-arabinofuranosidase from purple sweet potato were performed on the 40, 50, 60 and 700C; and 150, 180, 210 minutes. The extraction was done by ethanol solvent of 60,32% (v/v) acidified with citric acid of 2,39% (b/v). The measurement of anthocyanin levels using UV-Vis Spectrophotometer at 527 nm and 700 nm wavelengths at pH 1,0 and 4,5. The optimum condition of non-enzyme-assisted extraction was at 600C for 210 minutes, with the anthocyanin levels of 26,3842 mg/L; while with the AbfA enzyme-assisted at 500C for 180 minutes, with the anthocyanin levels of 28,2056 mg/L. The extraction with enzyme-assisted resulted the anthocyanin levels of 6,90% higher than without the using of enzyme.
ISOLASI DNA METAGENOMIK DALAM RANGKA STUDI METANOGENESIS PADA TANAH SAWAH Ni Luh Putu Mustika Praptiwi; Iryanti Eka Suprihatin; I Nengah Wirajana
Jurnal Kimia (Journal of Chemistry) Vol. 8, No. 1 Januari 2014
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (133.292 KB) | DOI: 10.24843/JCHEM.2014.v08.i01.p08

Abstract

Metagenomic is a new approach of complex genomes analysis from environmental samples. Isolation of total DNA from rice field soil sample has been conducted. Rice field, as an agriculture land, has been known to contribute 76% of total methane (CH4). The purpose of this study was to obtain metagenomic DNA isolated from rice field soil in order to study methanogenesis. Metagenomic DNA was isolated by direct and indirect cell lysis methods. Direct cell lysis method was conducted by directly lysing the cells in soil matrix followed by separation of DNA from the matrix and cell debris. Isolation of metagenomic DNA with indirect cell lysis method was carried out by cell separation from soil matrix followed by cell lysis. Metagenomic DNA were analyzed by agarose gel electrophoresis and UV-Vis spectrophotometry at ? 230, 260, and 280 nm. The results showed that metagenomic DNA could be isolated from rice field soil samples using direct and indirect cell lysis methods. Electrophoresis results showed that total DNA quality obtained by the indirect cell lysis was relatively less fragmented compared with direct cell lysis method. Spectrophotometric analysis showed that the total DNA isolated by indirect cell lysis was contaminated by humic acid more than metagenomic DNA isolated by direct cell lysis method. However, the metagenomic DNA by indirect cell lysis was contaminated by protein less than metagenomic DNA that obtained by direct cell lysis method.
PENENTUAN INDUSER MEDIA PERTUMBUHAN, SUHU DAN KONSENTRASI ION KALSIUM OPTIMUM LIPASE DARI MIKROBA LIPOLITIK TANAH HUTAN MANGROVE PANTAI SUWUNG KAUH BALI I P. J. D. A. Suartama; I N. Wirajana; A. A. B. Putra
Jurnal Kimia (Journal of Chemistry) Vol. 15, No.1, Januari 2021
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/JCHEM.2021.v15.i01.p16

Abstract

Lipase memainkan peranan penting dalam era industri green chemistry yang perlu dieksplorasi dari berbagai sumber untuk mengetahui kondisi optimum yang dibutuhkan agar diperoleh aktivitas enzim tertinggi. Tujuan penelitian ini adalah untuk menentukan jenis induser dalam media pertumbuhan, suhu dan konsentrasi ion kalsium (Ca2+) optimum lipase dari mikroba lipolitik tanah hutan mangrove pantai Suwung Kauh Bali. Mikroba lipolitik yang digunakan pada penelitian ini adalah mikroba lipolitik tunggal (ML.THM1) dan konsorsium mikroba lipolitik (KML.THM1) yang telah diisolasi dari tanah hutan mangrove. Pantai Suwung Kauh Bali pada penelitian sebelumnya. Mikroba lipolitik ditumbuhkan pada 3 (tiga) macam komposisi media yang berbeda dalam hal penambahan induser yaitu minyak zaitun (MSL.A), minyak jelantah (MSL.B) dan tanpa penambahan induser (MSL.C). Aktivitas lipase ditentukan dengan menggunakan metode titrasi asam basa. Jenis induser media pertumbuhan mikroba tunggal lipolitik dan konsorsium mikroba lipolitik yang optimum adalah minyak zaitun (MSL.A). Suhu optimum lipase dari konsorsium mikroba lipolitik tanah hutan mangrove (KML.THM1) adalah 37oC dengan aktivitas lipase sebesar 0,0667 ± 72,95 x 10-4 U/mL. Konsentrasi ion kalsium (Ca2+) optimum lipase dari konsorsium mikroba lipolitik (KML.THM1) pada suhu 37oC, 39oC, dan 41oC adalah 15 mM. Kata Kunci: ion kalsium, konsorsium mikroba, lipase, lipolitik, tanah hutan mangrove.
PEMURNIAN AMILASE MIKROBA AMILOLITIK DENGAN FRAKSINASI AMONIUM SULFAT DAN AMOBILISASI PADA AGAR-AGAR KOMERSIAL I N. Wirajana; R. R. Sirait; P. Suarya
Jurnal Kimia (Journal of Chemistry) Vol. 15, No.1, Januari 2021
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/JCHEM.2021.v15.i01.p07

Abstract

Peningkatan penggunaan biokatalis amilase membutuhkan pemurnian dan amobilisasi enzim ini untuk berbagai keperluan yang lebih ekonomis. Tujuan penelitian ini adalah menentukan persen kejenuhan amonium sulfat untuk pemurnian amilase mikroba amilolitik dan persen konsentrasi agar-agar komersial terbaik untuk mendapatkan persen efisiensi dan kestabilan tertinggi.Amilase diproduksi dari isolat mikroba amilolitik dengan kode UU1.1. Ekstrak kasar amilase ekstraseluler difraksinasi dengan amonium sulfat dengan tingkat kejenuhan 0-20%, 20-40%, 40-60%, 60-80% dan 80-100%; selanjutnya tiap fraksi dilakukan dialisis dalam buffer fosfat pH 6. Pengukuran aktivitas amilase dilakukan dengan menentukan kandungan gula pereduksi sebelum dan setelah reaksi enzimatis yang diinkubasi pada suhu 37oC pH 6 selama 60 menit dengan metode Dinitrosalicylic acid (DNS). Penentuan kadar protein total setiap fraksi diukur dengan metode Biuret. Aktivitas spesifik amilase ditentukan dari hasil pembagian aktivitas amilase dengan kadar protein total setiap fraksi. Amobilisasi dilakukan pada konsentrasi agar-agar 1%, 2% dan 3% (b/v). Penentuan persen agar-agar terbaik untuk amobilisasi amilase ditentukan dari efisiensi amilase teramobil tertinggi dan kestabilannya. Tingkat kejenuhan amonium sulfat 20-40% atau fraksi 2 diperoleh aktivitas spesifik amilase tertinggi sebesar 6,0 U/mg, yang merupakan tingkat kemurnian amilase tertinggi. Aktivitas amilase tertinggi sebesar3,3 x 10-3 U/mL, diperoleh dari hasil fraksinasi pada tingkat kejenuhan amonium sulfat 40-60% atau fraksi 3, digunakan untuk amobilisasi dalam matriks agar-agar komersial. Amobilisasi amilase dengan efisiensi dan kestabilan tertinggi diperoleh pada konsentrasi agar-agar 3% (b/v), baik untuk ekstrak kasar amilase maupun amilase hasil fraksinasi. Kata kunci: agar-agar, amilase, amilum, amobilisasi, fraksinasi. Increased use of amylase biocatalysts requires the purification and immobilization of this enzyme for a variety of more economical purposes. The purpose of this study was to determine the percent saturation of ammonium sulfate for the purification of amylolytic microbial amylase and the best percent of commercial agar concentration to obtain the highest percent efficiency and stability. Amylase is produced from amylolytic microbial isolates with the code UU.1.1. Crude extract of extracellular amylase is fractionated with ammonium sulfate with saturation levels of 0-20%, 20-40%, 40-60%, 60-80% and 80-100%; then each fraction was dialyzed in a phosphate buffer 6. The measurement of amylase activity was carried out by determining the reducing sugar content before and after the enzymatic reaction incubated at 37oC pH 6 for 60 minutes with the Dinitrosalicylic acid (DNS) method. Determination of total protein content of each fraction was measured by the Biuret method. The specific activity of amylase is determined from the results of the division of amylase activity by the total protein content of each fraction. Immobilization is carried out at 1%, 2% and 3% (w / v) agar concentrations. Determination of the best agar agar for amylase immobilization is determined from the highest immobilized amylase efficiency and its stability. Ammonium sulfate saturation level of 20-40% or fraction 2 obtained the highest specific amylase activity of 6.0 U / mg, which is the highest level of purity of amylase. The highest amylase activity of 3.3 x 10-3 U / mL, obtained from fractionation at 40-60% ammonium sulfate saturation level or fraction 3, was used for immobilization in the commercial agar matrix. Amylase immobilization with the highest efficiency and stability was obtained at a concentration of agar 3% (w / v), both for crude extracts of amylase and fractionated amylase. Keywords: agar-agar, amylase, fractionation, immobilization, starch.
ISOLASI DNA METAGENOMIK DARI TANAH HUTAN MANGROVE PANTAI SUWUNG BALI I Nengah Wirajana; Darma Asih Yuliana; Ketut Ratnayani
Jurnal Kimia (Journal of Chemistry) Vol. 7, No. 1 Januari 2013
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (161.666 KB) | DOI: 10.24843/JCHEM.2013.v07.i01.p03

Abstract

Metagenomic DNA isolation from mangrove forest soils of Suwung Beach-Bali was conducted to exploit the biocatalytic potential of microbial communities for the discovery of novel cellulase. The whole DNA isolation was conducted by modification of preparation step by Marco (2010) and DNA extraction step by Amorim et al (2008). The results of metagenomic DNA isolation were analyzed by spectrophotometry and agarose gel electrophoresis. The results showed that whole DNA was able be isolated successfully, but protein and humic acid were found as contaminant.
Co-Authors A. A. B. Putra Anak Agung Istri Agung Mayun Laksmiwati D. Rizkiyanti Darma Asih Yuliana Dewi Andayani Farmawati Dewi, I Gusti Ayu Agung Gangga Samala Diah Suci Eddy Bagus Wasito Gusti A Malelak H.M. Soekry Erfan Kusuma I Gede Mahardika I Gusti Ngurah Bagus Andre Hartawan I Ketut Gede Dharma Dewantara I Made Agus Gelgel Wirasuta I P. J. D. A. Suartama I Putu Mahendra I W BUDIARSA SUYASA I Wayan Gede Gunawan I Wayan Suarsa I. A. Gede Widihati Ida Ayu Ratih Dwi Nugraha Putri Ida Bagus Amertha Putra Manuaba Ida Bagus Putra Manuaba Ida Bagus Putu Eristya Putra Ika Kurniawati Indra Juana Adikara Inten Hardianti Nizar Iryanti Eka Suprihatin James Sibarani Kazuo Sakka Ketut Ratnayani Khisan Qamariya Kimura, Tetsuya Kusuma, Soekry Erfan Luh De Dwi Jayanthi Luk Ketut Budi Maitriani M. Manurung Made Arsa Made Dharmesti Wijaya Made Rai Dwitya Wiradiputra Mirawati N.K.W. N. M. T. Juliasari N. W. Bogoriani Ni Komang Ariati Ni Komang Lia Wahyuni Ni Luh Md. Widayantini Ni Luh Putu Mustika Praptiwi Ni Made Suaniti Ni Made Yustikarini Ni Nengah Kartini Asih Ni Nyoman Tri Puspaningsih Ni Putu Ariantari Ni Putu Citra Anggryni Sugitha Ni Putu Frida Oktaningtias Widiarthi Ni Putu Rahayu Artini O. Ratnayani Oka Ratnayani P. Suarya Pradnyaniti, D.G Putra, Pramana Kumala Putri, Ni Wayan Prasanthi Swarna Putu Elvira Yulianthi Putu Suarya R. R. Sirait Ro’yal Aini Sagun Chandra Yowani Sakka, Kazuo Satriya Putra Prakoso Soekry Erfan Kusuma Tetsuya Kimura Wahyu Dwijani Sulihingtyas Y. H. Pratiwi