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All Journal Jurnal Pengolahan Hasil Perikanan Indonesia Forum Pasca Sarjana JOURNAL OF COASTAL DEVELOPMENT Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology
Ekowati Chasanah
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Education Environmental Science Immunology & microbiology

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Journal : JOURNAL%20OF%20COASTAL%20DEVELOPMENT

 1 
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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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. 
OPTIMIZATION OF Bacillus sp. K29-14 CHITINASE PRODUCTION USING MARINE CRUSTACEAN WASTE Agustinus Robert Uria; Ekowati Chasanah; Yusro Nuri Fawzya
JOURNAL OF COASTAL DEVELOPMENT Vol 8, No 3 (2005): Volume 8, Number 3, Year 2005
Publisher : JOURNAL OF COASTAL DEVELOPMENT

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Abstract

Chitin is present in large quantities in the marine crustacean waste disposed by seafood processing industries, making it very desirable as the substrate for producing chitinase, a hydrolytic enzyme of considerable interest in many industrial and agricultural applications. In our work, crustacean waste powder and its combination with colloidal chitin at different concentrations (0.5, 1.0, and 1.5%) were utilized to optimize the chitinase production by the bacterium, Bacillus sp. K29-14. The results showed that the chitinase production with the three different substrate concentrations was relatively constant in the range of 0.2 to 0.3 U/ml during 12 days cultivation, although there was a bit reduction after day 8. This activity profile seems to be similar to that of the protein content. Whereas the chitinase production on the media containing crustacean waste powder and its combination with colloidal chitin at the three concentrations showed that the highest activity (3.0 to 4.6 U/ml) was achieved on day 7 and 8. The specific chitinase activity with the waste powder at different concentrations of substrate (0.5, 1.0 and 1.5%) was increasing slowly during a nine-day cultivation. The optimal chitinase production (4.6 U/ml) was achieved with the combined substrate of 0.5% on day 8.
MARINE BIODISCOVERY RESEARCH IN INDONESIA : CHALLENGES AND REWARDS Ekowati Chasanah
JOURNAL OF COASTAL DEVELOPMENT Vol 12, No 1 (2008): Volume 12, Number 1, Year 2008
Publisher : JOURNAL OF COASTAL DEVELOPMENT

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Abstract

Marine biodiscovery or bioprospecting activity is a search for marine products derived from marine biodiversity that can be developed for various industrial needs. Including in this activity is the process of identifying chemical compounds made by biological organisms which is often called natural product discovery. Indonesia, well known as a mega-diversity country, is one of the world hot sport of marine biodiversity. The richness of biodiversity is claimed as mirror of the richness of the chemical compounds, therefore, Indonesian waters might be rewarded with variety of chemical compounds thought to be an endless source of novel drugs and drug leads for pharmaceutical use. Up to 2007, at least 77 new compounds from 14 sponges and 19 new compounds from non-sponge organisms with pharmacological potential have been identified from Indonesian waters. To make this richness potentials becoming real in economic value, many factors should be considered. The bioactive is produced in small quantity, and the lengthy process from discovery step of a novel compound to the preclinical and clinical trials step is usually becoming a problem. Mari culture might be one among methods that can be developed in Indonesia to overcome the degradation hazard of marine resources. Conducive environment for investments, and improvement of technology on marine bioactive production through mariculture are factors to be improved to initiate and develop a sustainable biotechnology industries in Indonesia. 
NOVEL MOLECULAR METHODS FOR DISCOVERY AND ENGINEERING OF BIOCATALYSTS FROM UNCULTURED MARINE MICROORGANISMS Agustinus Robert Uria; Yusro Nuri Fawzya; Ekowati Chasanah
JOURNAL OF COASTAL DEVELOPMENT Vol 8, No 2 (2005): Volume 8, Number 2, Year 2005
Publisher : JOURNAL OF COASTAL DEVELOPMENT

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Abstract

Metagenomics is a powerful cultivation-independent approach, which can be applied to gain access to the biocatalysts from uncultured marine microorganisms. Discovery of marine biocatalysts by this approach, in general, involves four main steps. First, a metagenomic library containing a pool of biocatalyst-encoding genes is constructed from a marine environment, which can be done by various methods, including cloning of enzymatically-digested DNA, uncut DNA, and PCR-amplified products. Second, the metagenomic library is screened for the genes of interest by employing the activity assay of expression product, in situ  hybridization, or Polymerase Chain Reaction (PCR). Third, the obtained target genes, both functional and phylogenetic genes, are sequenced and analysed by using bioinformatic tools in order to gain information on the functional and structural properties as well as the microbial sources of the encoded biocatalysts. Finally, the target genes are expressed in suitable microbial hosts, thereby producing the corresponding recombinant biocatalysts. All existing methods in engineering of marine biocatalysts for the performance improvement can be classified into two main strategies: (i) rational design and (ii) directed evolution. Rational design, which may include the use of resctriction enzyme(s) and splicing by overlap extension (SOE), requires information on the biocatalyst`s structural and functional properties to alter specific amino acid(s). Whereas directed evolution, including error-prone PCR technique and gene shuffling, needs no such information.
Co-Authors Agustinus R Uria Agustinus Robert Uria Andi Hakim Anthony D. Wright Catherine H. Liptrot Cherie A. Motti DAHRUL SYAH Dewi Seswita Zilda Dianne M. Tapiolas Gintung Patantis Hedi Indra Januar Hedi Indra Januar Ifah Munifah Ikhsan B. Waluyo Joko Santoso Kustiariyah Tarman M.R.R. Lukie Trianawati MAGGY T. SUHARTONO Mahrus Ali Muhammad Nursid Muhammad Nursid Nurrahmi Dewi Fajarningsih Putalan, Reinal Tanti Yulianti TATI NURHAYATI Uju Uju Usman Sumo Friend Tambunan Yenny Risjani Yusro Nuri Fawzya Yusro Nuri Fawzya