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PURIFICATION AND CHARACTERIZATION OF Aeromonas media KLU 11.16 CHITOSANASE ISOLATED FROM SHRIMP WASTE Ekowati Chasanah; Gintung Patantis; Dewi Seswita Zilda; Mahrus Ali; Yenny Risjani
JOURNAL OF COASTAL DEVELOPMENT Vol 15, No 1 (2011): Volume 15, Number 1, Year 2011
Publisher : JOURNAL OF COASTAL DEVELOPMENT

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Abstract

Our previous study found that KLU 11.16, isolated from shrimp waste secreted chitinolytic enzymes. The crude enzyme was interesting since their chitooligosccharide was able to inhibit some pathogenic bacteria. In this study we report a purification and characterization of the chitosanase enzyme produced and the identification of the KLU 11.16. Purification of the enzyme was done two steps by ion exchange chromatography followed by gel filtration. Two out of 4 peaks from Gel Filtration step, i.e. fraction 16 and 33 were capable of hydrolyzing 100% deacetylated chitosan, indicating that both fractions contained chitosanase enzyme. The enzyme from fraction 16 had approximate molecular weight of 98.3 kDa. The enzyme worked optimally at temperature of 300C, and pH 6. Addition of Ca2+, Fe2+, K+, Na+ ions in the form of Cl2 salt and detergent Triton X-100 increased the enzyme activity, while Co2+, Mn2+ and Zn2+ ions in the same concentration decreased the enzyme acitivity. Addition of EDTA and SDS significantly decreased the enzyme activity. Molecular based identification revealed that KLU 11.16 was 99% similar to Aeromonas media.
SCREENING AND CHARACTERIZATION OF BACTERIAL CHITOSANASE FROM MARINE ENVIRONMENT Ekowati Chasanah; Dewi Seswita Zilda; Agustinus R Uria
JOURNAL OF COASTAL DEVELOPMENT Vol 12, No 2 (2009): Volume 12, Number 2, Year 2009
Publisher : JOURNAL OF COASTAL DEVELOPMENT

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Abstract

Screening of extracellular chitosanase from bacterial isolates associated with marine sponges have been done. Out of 100 bacterial isolates, forty isolates were capable of forming clearing zones on the chitin media and one isolate, 34-b, produced the highest chitinolytic index. The enzymes was produced on chitin liquid medium at 37oC in a shaking waterbath for a five-day cultivation. Crude enzymes were prepared by cell-free supernatant (CFS) and concentrated through 70% (saturated) ammonium sulphate percipitation followed by dialysis. The enzymes worked best at pH and temperature of 6-7 and 60oC, respectively. The half-life (T1/2) for chitosanase activity was 500.2 min or 8.34 hours (at 37oC) and 55.12 min (at 50oC), indicating the enzyme are quite stable at that temperature. However, around 80% of the original activity was lost at 60oC after 15 min of incubation. 
Teknik Peningkatan Performa Enzim Asal Mikroba Laut yang Tidak dapat dikultur Agustinus Robert Uria; Dewi Seswita Zilda; Yusro Nuri Fawzya
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 1, No 1 (2006): December 2006
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v1i1.72

Abstract

Tingginya keanekaragaman dan kelimpahan mikroba laut menunjukkan potensinya sebagai sumber paling menjanjikan untuk penemuan produk alami yang bernilai industri.  Di antara berbagai bentuk produk alami, enzim termasuk paling diminati dalam dunia industri baik dalam industri kimia, farmasi maupun makanan.  Hal ini berkenaan dengan peranannya sebagai katalis yang ramah lingkungan, ekonomis dan bersih (Wahler Reymond, 2011).
Cloning of a Transglutaminase Gene from Streptomyces thioluteus TTA 02 SDS 14 Seprianto Seprianto; Dewi Seswita Zilda; Yusro Nuri Fawzya; Suharsono Suharsono; Puspita Lisdiyanti; Agustinus Robert Uria
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 11, No 1 (2016): May 2016
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v11i1.189

Abstract

Microbial Transglutaminase (MTGase, EC 2.3.2.13) is an enzyme that catalyzes the transfer of acyl group. Many microbial strains that produce MTGase belong to Streptomyces members. This research was aimed at cloning of a MTGase gene. PCR–based screening of ten MTGase-producing streptomyces isolates from soil in West Nusa Tenggara led to detection of one potential isolate, designated as TTA 02 SDS 14. The partial  MTGase-encoding gene (470 bp)  was amplified by PCR and sequenced. The sequence result indicate its similarity of 93 % with that of Streptomyces cinnamoneus. The 16S rRNA gene analysis showed its identity as Streptomyces thioleteus. Fosmid-based construction of a genomic library from the isolate  and subsequent screening led to the isolation of  a ~40-Kb fosmid harboring a MTGase gene.
SCREENING OF THERMOSTABLE PROTEASE PRODUCING MICROORGANISMS ISOLATED FROM INDONESIAN HOTSPRING Dewi Seswita Zilda; Eni Harmayani; Jaka Widada; Widya Asmara; Hari Eko Irianto; Gintung Patantis; Yusro Nuri Fawzya
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 7, No 3 (2012): December 2012
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v7i3.5

Abstract

Although many proteases had been studied and characterized, only a few of them are commercially available.  Protease thermostability is one of the crucial properties for industrialapplication. This research aimed to isolate and to screen the potential isolate which produce thermostable protease. There were 6 isolates (BII-1, BII-2, BII-3, BII-4, BII-6 and LII), isolated using solid Minimal Synthetic Medium (MSM) supplemented with 1.5% skim milk, that have, protease activity. Based on the 16S-rRNA gene sequencing analysis, isolates BII-1, BII-2 and BII- 6 were identified as Bacillus licheniformis, isolates BII-3 and BII-4 were identified as Bacillus subtilis, while isolate LII was identified as Brevibacillus thermoruber. Three isolates (BII-6, BII-4 and LII) were then further investigated for the second screening step using liquid MSM supplemented with 1% skim milk. The isolates (BII-6, BII-4 and LII) optimally produced protease when they were cultivated at 35, 30 and 50o C respectively after 22 h of incubation. Protease produced by BII-6, BII-4 and LII had optimum temperature  of 65, 60 and 85o C, optimum pH at 7-8, 8 and 9 and stable up to 100 min at 55, 60 and 75o C respectively.
BACTERIAL DIVERSITY OF THE DEEP SEA OF SANGIHE TALAUD, SULAWESI Gintung Patantis; Ekowati Chasanah; Dewi Seswita Zilda; Ikhsan B. Waluyo
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 7, No 1 (2012): May 2012
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v7i1.12

Abstract

Deep sea is an extreme environment characterized by cold temperature, high pressure, lackof  light and nutrients. Microorganisms live in these habitat are unique microorganisms andknown to have tremendous source of potential agents for biotechnology processes. Indonesia asan archipelagic country has a vast deep ocean. This study aims to see the diversity of bacteria inSangihe Talaud Deep Sea, Sulawesi. Analysis of bacterial diversity was carried out by culturedand uncultured method. Terminal Restriction Fragment Length Polymorphism (T-RFLP) techniquewas used for uncultured analysis of the microorganisms biodiversity, while cultured one wasdone by plating the samples of water onto Zobell media. The results showed that, there were 21isolates obtained by cultured method. The identification which based on 16S rDNA by PCR methodshowed the genus of Pseudomonas, Pseudoalteromonas, Alteromonas, Vibrio, Shewanella andUncultured bacterium were identified. However, 14 classes of bacteria were obtained by usingTRFLP method i.e Acetobacteraceae class, Actinobacteria, α-proteobacteria, -proteobacteria, δ-proteobacteria, γ-proteobacteria, Bacili, Bacteroidetes, Chlorobi, Chroococcales, Clostridia,Erysipelotrichi, Synergistia, and Zetaproteobacteria. here were also  unclassified bacteria anduncultured bacterium found in the samples.
Purification and Characterization of Glucose Oxidase of Aspergillus niger IPBCC.08.610 ANINDA INDRIANI; LAKSMI AMBARSARI; DEWI SESWITA ZILDA
Microbiology Indonesia Vol. 12 No. 2 (2018): June 2018
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (575.093 KB) | DOI: 10.5454/mi.12.2.2

Abstract

Glucose oxidase was an enzyme which catalyzed β-D-Glucose to gluconic acid and hydrogen peroxide. Glucose oxidase from Aspergillus niger IPBCC.08.610 was isolated, purified and characterized. The enzyme was purified by ammonium sulphate precipitation and dialysis. The specific activity and yield of dialysis fraction were 19.766 U/mg and 4.744%. The optimum pH and temperature were 6 and 30oC. The stability of enzyme at optimum pH and temperature was decreasing 50% at 25 minutes. The km and vmax values for enzyme were 27 mM and 0.986 U/mg.