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All Journal JOURNAL OF COASTAL DEVELOPMENT Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology
Agustinus Robert Uria
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Education Environmental Science Immunology & microbiology

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Journal : JOURNAL OF COASTAL DEVELOPMENT

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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.
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 Dewi Seswita Zilda Dewi Seswita Zilda Ekowati Chasanah Irvina Nurrachmi Kusnadi, Yudi Priyono, Franciscus Edi PUSPITA LISDIYANTI Seprianto, Seprianto Suharsono Suharsono Tri A Hadi Yusro Nuri Fawzya Yusro Nuri Fawzya