A.R. Uria,
I. Munifah, and E. Chasanah, 2007. Isolation and Enzymatic
Fragmentation of Genomic DNA from Uncultured Microbial Symbionts of
Sponge as Prerequisite for Metagenomic Library Construction. Indonesian Fisheries Research Journal 13(2): 125-130. Uria A.R., D.S. Zilda, dan Y.N. Fawzya, 2006. Teknik Meningkatkan Performa Enzim Asal Mikroba Laut yang Tidak Dapat Dikultur. SQUALEN: Bulletin Pascapanen & Bioteknologi Kelautan Dan Perikanan 1(1) 2006 : 7-14. (for the detail, please contact A.R. Uria at uria_biotek@yahoo.com) Uria, A.R., Y.N. Fawzya, and E. Chasanah, 2005. Novel Molecular Methods for Discovery and Engineering of Biocatalysts from Uncultured Marine Microorganisms. J. Coastal Development 8(2):53-74.
Abstract: Biocatalysts originated from marine microorganisms have attracted much scientific attention for being used in various industrial processes. However, for many years the search for new biocatalysts of microbial origin was based on cultivation-dependent methods which count only less than 1% of all microorganisms in most environments, including marine environment. However, a recent breakthrough in Microbial Ecology, marked with the birth of Metagenomics, has allowed to gain access to the enormous uncultured microbial diversity for the discovery of an unlimited pool of new biocatalysts. Metagenomic approach for the discovery of biocatalysts from uncultured marine microorganisms, in principle, involves (i) construction of metagenomic library, (iii) screening of metagenomic library, (iii) sequence analysis of genes, and (iv) expression of genes of interest in appropriate hosts. With recent advances in DNA sequencing technology and bioinformatics tools, this approach have expanded into the sequence-based analysis of biocatalysts-encoding genes and phylogenetic markers. The analysis results can give useful information on the functional and structural properties and microbial source of the target biocatalysts. In spite of the potential of finding new biocatalyst from marine microorganisms, it often appears that natural biocatalysts need to be improved in order to meet industrial and biotechnological requirements. Improving the biocatalyst performance at molecular level, which is popularly called protein engineering, can be classified into two main categories: rational design and laboratory evolution. Rational design needs the information on the structure of protein or enzyme as well as the relationship between sequence, structure and function. This information is not required for employing laboratory evolution.Uria, A.R., Y.N. Fawzya, and E. Chasanah, 2005. Eksplorasi Enzim Mikroba dari Lingkungan Laut melalui Pendekatan Metagenomika (Exploration on Microbial Enzymes from Marine Environment using Metagenomic Approach). A review paper in Indonesia language. WPPI 11(7):17-24.
Abstract: Selama bertahun-tahun pencarian enzim baru dari mikroba didasarkan pada isolasi dan pembiakkan mikroba di laboratorium (culturable microbes). Akibatnya, sebagian besar enzim yang diterapkan sekarang hanya berasal dari culturable microbes. Padahal culturable microbes hanya mencakup sekitar 1% dari semua mikroorganisme yang hidup di alam, termasuk di lingkungan laut. Sedangkan unculturable microbes yang prosentasenya sekitar 99% sejauh ini belum banyak diungkap. Terobosan-terobosan yang spektakuler dalam bioteknologi dewasa ini telah mendorong lahirnya pendekatan baru untuk mengakses enzim baru dari unculturable microbes. Pendekatan yang dikenal dengan sebutan Metagenomika ini melibatkan (i) pengumpulan gen-gen dari laut, (ii) penyeleksian gen-gen yang diinginkan, (iii) analisis gen target, dan (iv) ekspresi gen target. Khususnya, hasil analisis gen target memberikan gambaran tentang sifat-sifat fungsional dan struktural dari enzim yang disandinya. Sedangkan hasil analisis gen filogenetik (seperti gen 16S rRNA) dapat menunjukkan asal-usul enzim yang disandi oleh suatu gen target. (click here for the full text) | |
