KARAKTERISASI ENZIM AMILASE DARI ISOLAT BAKTERI TERMOFILIK BACILLUS SUBSTILIS

Dalam penelitian ini digunakan isolat bakteri termofilik yang diisolasi dari sumber air panas Makula Tana Toraja. Isolasi enzim dilakukan setelah bakteri tersebut diaktifkan dan dikultur dalam medium yang mengandung pati pada suhu 500C, pH 7 selama 72 jam. Enzim amilase ekstraseluler yang diperoleh memiliki keaktifan optimum pada suhu 35oC dan pH 6,4. Enzim ini merupakan enzim amilase logam karena keajtifan katalitiknya dapat ditingkatkan oleh ion logam, seperti Co2 +, Mg 2 +, Ni 2 +, and Ca2 + sehingga bersifat aktivator untuk ion Zn 2 + menurunkan aktivitas enzim dan bersifat inhibitor. Ekstrak kasar enzim amilase hasil isolasi dapat menghidrolisis masing-masing substrat yaitu pati sagu, pati singkong dan pati jagung sebesar 92,36 %, 83,15 % dan 61,57 %. Aktivitas spesifik enzim amilase pada kondisi optimum\ud diperoleh 0.00142 U/mg protein

Patong Proceeding of The International Seminar on Chemistry

Abstract To better understanding of DNA replicating-coupled chromatin assembly and transcription regulation, we cloned and sequenced cDNA encoding the chicken p46 polypeptide, chp46, homologous to the p48 subunit of chicken chromatin assembly factor-1, chCAF-1p48. The cDNA encoding a protein consists of 424 amino acids including a putative initiation Met, is a member of the WD protein family, with seven WD repeat motifs, and exhibits 90.3% identity to chCAF-1p48, and 94.3% identity to the human and mouse p46 polypeptides. The p46 polypeptide fusion protein were synthesized by in vitro translation system and expressed in Escherichia coli under induction by 50 µM IPTG and single step purified with glutathione-agarose beads, showed that GST-tagged protein of approximately 72 kDa, were dramatically synthesis in Escherichia coli BL-21 cells. The in vitro experiment established that chp46 interacts with chicken histones, chHDAC-1, and chHAT-1. The in vitro immunoprecipitation experiment, involving truncated mutants of chp46, revealed not only that two regions comprising amino acids 33-179 and 375-404 are necessary for its binding to H2B, but also that two regions comprising amino acids 1-32 and 405-424 are necessary for its binding to H4. Furthermore, the GST pulldown affinity assay, involving truncated mutants of chp46, revealed that a region comprising amino acids 359-404 binds to chHAT-1 in vitro. Taken together, these results indicate not only that chp46 should participate differentially in a number of DNA-utilizing processes through interactions of its distinct regions with histones and chHAT-1, but also that the proper propeller structure of chp46 is not necessary for its interaction with chHAT-1

PENGARUH SUBSTRAT DAN ION LOGAM TERHADAP AKTIVITAS ENZIM LIPASE DARI Aspergillus oryzae PADA KOPRA BERJAMUR

Penelitian ini menggunakan isolat fungi Aspergillus oryzae dari kopra ber-jamur sebagai sumber enzim lipase. Isolasi enzim dilakukan setelah fungi tersebut diaktifkan dan dikultur pada masing-masing media fermentasi yang mengandung substrat olive oil, virgin oil, triolein dan palm oil pada suhu 370C dan pH 7,0 selama 8 hari. Enzim lipase yang dihasilkan dilakukan uji aktivitas, dan dari uji aktivitas didapatkan bahwa enzim lipase dapat menghidrolisis semua substrat, dan aktivitas paling tinggi diperoleh pada substrat triolein. Uji aktivitas dilakukan dengan menam-bahkan ion-ion logam yaitu: Co2+, Ca2+, Mg2+, Ni2+, Mn2+, Zn2+ dan EDTA, dengan konsentrasi masing-masing 1mM dan 10 mM. Dari hasil penelitian diperoleh ion-ion Co2+, Ca2+ dan Mg2+ dapat meningkatkan aktivitas enzim sebesar masing-masing Co2+ = 19% dan 6%, Ca2+ = 4% dan 8% pada konsentrasi 1mM dan 10 mM, Mg2+ tidak meningkatkan aktivitas enzim pada konsentrasi 1mM, tetapi pada konsentrasi 10 mM meningkatkan aktivitas enzim 11%. Ion Ni2+, Mn2+ dan Zn2+ tidak meningkatkan aktivitas enzim, demikian pula dengan penambahan EDTA

PEMURNIAN DAN KARAKTERISASI ENZIM LIPASE DARI ASPERGILLUS ORYZAE PADA KOPRA BERJAMUR

An investigation on purification and characterization of lipase enzyme production Aspergillus oryzae from copra by fermentation of olive oil has been carried out. This enzyme can be produced by fermenting olive oil in a medium containing Aspergillus oryzae. Crude enzyme is obtained by centrifuging the medium cultures containing Aspergillus oryzae at 3500 rpm for 30 minutes and than adding 0,2 M borat buffer (pH 8,2). Enzyme activity was determined from paranitrophenol as product of lipase catalysis of paranitrophenylbutirat (0,2 M) as substrate measured by the Vorderwulbecke method. Prepurification process was by ammonium sulphate fractionation. Precipitation by and 60-80% ammonium sulphate produced maximum activity of enzyme. Purification by Q sepharosa FF and sephadex G-75 collum chromatography produced four and three fractions with purifity of 12,85 and 20,25 times than crude enzyme respectively. Characterization of this enzyme showed optimum condition at pH 8,2, temperature at 350C, the Km value at 0,046 M, and Vmaks is 1,926 ??mol/menit and the melecular weight at 40,7 kDa

ANALISIS KADAR LOGAM Cu DALAM BAKTERI SIMBION PADA SPONS Callyspongia sp

This research aims to analyze the levels of copper (Cu) in the sponge symbiont bacterial. The research was carried out through the steps of isolation, identification, and measurement of the metal content. Sponges used in this research is of Callyspongia sp. Bacteria were isolated from Callyspongia sp sponges in the Bacillus sp gram-positive. By using Atomic Absorption Spectrophotometer, the results showed that the levels of Cu for Callyspongia sp is 8.9112 ppm, while in the extracellular bacterial isolates Cu level ranging between 1.4009 ppm and 2.0336 ppm and in the intracellular is between 0.4786 ppm and 1.7813 ppm. It is concluded that bacterial isolates either in extra- as well as intracellular were both accomodated copper in their bodies

IMMOBILIZATION OF GLUCOSE OXIDASE ENZYME FROM Penicillium sp-3 LOCAL STRAIN

Immobilization of glucose oxidase (GOD) enzyme from Penicillium sp-3 local strain has been carried out using ionotropically entrapping method in  Ca-alginate membrane coupled with Na-polyacrilate. The entrapment of the enzyme in diffusion membrane occur spontaneously by cross-linking between Na-alginate/Na-polyacrilate and CaCl2. The GOD enzyme immobilized by addition of 1 % Na-polyacrilate has the highest encapsulation efficiency, that is 87.13 % with the smallest percentage of diffusion, i.e. 23.37% and the relative activity of 50%. The GOD immobilized enzyme had good stability at the pH range 4 - 7 during 30 minutes of storage and was stable at a temperature of 20 oC. The activity of the GOD enzyme after being utilized continuously for 5 times only decrease up to 47,06 % compared to that in the initial utilization.   Keywords: immobilization, glucose oxidase, Penicillium sp-3, calcium alginate, sodium  polyacrilate

Studi Bioaktivitas Antibakteri Fraksi Protein Hidroid Aglaophenia cupressina Dari Pulau Lae-Lae Sulawesi Selatan

This research aimed to know the activity of bioactive protein, extracted from Hydroid (Aglaophenia cuppressina), in inhibiting the growth of pathogenic bacteria. Samples were taken from Lae-lae Island in South Sulawesi. Methods used were the Lowry method for determining the protein concentration and the agar diffusion method  for testing the antibacterial activity. Extraction of Hydroid was conducted by making use of buffer solution, (0,1 M Tris-HCl of pH 8.3, 2 M NaCl, 0.01 M CaCl, 1% β-mercaptoethanol, and 0.5% Triton X-100). Protein was purified by ammonium sulfate precipitation at 20%, 40%, 60% and 80% saturation followed by dialysis. Results showed that the protein fraction of Aglaophenia cuppressina before dialysis with 40 % ammonium sulfate saturation had the highest antibacterial activity to Staphylococcus aureus and Escherichia coli with inhibition zones of  14.58 and 18.05 mm, respectively. In addition, the protein fraction of Aglaophenia cuppressina after dialysis with 40 % saturation also had the highest antibacterial activity to the tested bacteria with inhibition zones of  13.95 and 11.78 mm, respectively. The minimum inhibition concentration to the tested bacteria and Escherichia coli at 40% ammonium sulfate (0,1 M Tris-HCl of pH 8.3, 2 M NaCl, 0.01 M CaCl 2 saturation was 6000 µg/mL with inhibition zones of 9.78 and 8.73 mm, respectively. Based on the bioactivity test of the Hydroid protein fraction using the agar diffusion method, it can be concluded that the activity characteristic of the fraction is bacteriostatic.Key words: Bioactivity, hydroid, bioactive protein, antibacterial, inhibition zone

Isolation and Characterization of Cellulase from Mussel Shells Atactodea Striata Using Substartes of Paper Cellulose

Cellulase enzymes are hydrolitic enzyme capable c.f hydrolyzing cellulose to glucose monomers that can be isolated from mussel shells (Atacodea strain) using cellulose paper substrateThis study aims to determine the optimum condition for cellulase enzymes from mussel sheels covering the substrate concentration, pH, temperature, and concentratiom of the enzyme and determine the value of KM and VMax. This study begins by isolating cellulase enzymes through the cell destruction, homogenation, and centrifugation. Determination of optimum conditions can be seen from values of enzyme activity and determined by measuring glucose levels. Glucose levels were determiden based on obtained Nelson-Somogy method. Determintion of protein was 2%, the optimum temperature was 40 C, the optimum pH was 5.4 and the optimum enzyme concentration was 12 mg/mL, respectively. KM and VMax values was at 1.192% (w/v) and 1.12 umol/mL/min. At the optimum conditions, the specific activity of the cellulase enzyme from mussel sheels (Atactodea striata) in hydrolyzing cellulose contained in old newspapers into glucose was 1.62 x 10 -4 U/mg

Pemanfaatan Kitin Sebagai Bahan Membran Elektroda Enzim Diamin Oksidase Untuk Biosensor Histamin

This research aims to utilize isolated chitin from shrimp waste to develop histamine biosensors basedon diamine oxidase (DAO) enzyme electrode with cyclic voltammetry. DAO enzyme trapped in the chitin-cellulose acetate membranes with various comparisons were layered on the Pt electrode. Histamine will be oxidized by the DAO enzyme produces aldehydes and H2O2 which acts as an electron transfer mediator. Biosensor performance is influenced by several factors, especially the concentration and composition of electrode  membranei.Comparison of chitin-cellulose acetate used in this study were 1:1, 2:1 and 3:1. Isolated chitin from the shrimp waste is chemically obtainedrendamen of 23.6%, and characterization of electrode membrane by FTIR and cyclic voltammetry showed that the DAO enzymes electrode with chitin-cellulose acetate membrane 2:1 is the best composition

Pemanfaatan Kitin Sebagai Bahan Membran Elektroda Enzim Diamin Oksidase untuk Biosensor Histamin

Penelitianinibertujuanmemanfaatkan kitin yang diisolasi dari limbah udang untukmengembangkanbiosensor histamin berbasiselektroda enzimdiamin oksidase (DAO)denganvoltammetri siklik.EnzimDAOdiperangkapdalammembran kitin-selulosa asetatdengan berbagai perbandingan yangdilapiskanpadaelektrodaPt. Histamin akan dioksidasi oleh enzim DAO menghasilkan aldehid dan H2O2yangbertindaksebagaimediatortransferelektron. Kinerjabiosensorinidipengaruhiolehbeberapafaktor,terutama konsentrasi dan komposisi membran elektroda.Perbandingan kitin-selulosa asetat yang digunakan pada penelitian ini adalah 1:1, 2:1, dan 3:1. Hasil isolasi kitin dari limbah udang secara kimiadiperoleh rendamen sebesar 21,5%, dankarakterisasi dengan FTIR dan voltammetri siklik menunjukkan bahwa elektroda enzim DAO dengan membran kitin-selulosa asetat 2:1 merupakan komposisi terbai