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All Journal JOURNAL OF COASTAL DEVELOPMENT Microbiology Indonesia Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology
Dewi Seswita Zilda
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Education Environmental Science Immunology & microbiology Medicine & Pharmacology

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Journal : JOURNAL OF COASTAL DEVELOPMENT

 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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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. 
Co-Authors Agustinus R Uria Agustinus Robert Uria ANINDA INDRIANI Ekowati Chasanah Eni Harmayani Gintung Patantis Hari Eko Irianto Ikhsan B. Waluyo JAKA WIDADA LAKSMI AMBARSARI Mahrus Ali PUSPITA LISDIYANTI Seprianto, Seprianto Suharsono Suharsono Widya Asmara Yenny Risjani Yusro Nuri Fawzya